Proteomics Research 2013

A.

Phos-tag™ - Phosphorylated Protein Analysis

1. Phos-tag Acrylamide

2. SuperSep Phos-tag(Precast Phos-tag™ Acrylamide Gels)

3. Phos-tag Biotin

4. Phos-tag Mass Analytical Kit

5. Phos-tag Agarose

B.

Protein Electrophoresis

1. SuperSep - Precast Polyacrylamide Gels

2. EasySeparator™

3. Reagents for Gel Preparation

4. Protein Size Markers

5. Premixed Buffers · Reducing Agents

6. Staining Reagents

a. Negative Gel Stain MS Kit

b. Silver Staining Kits

c. CBB R-250 Stain Kits - Quick CBB -

C.

Western Blotting

1. Membranes for Blotting Applications

2. Blocking Reagents

3. Wash Solutions

4. Labeled 2nd Antibodies

5. Loading Control Antibodies

6. Antibody Detection Reagent

7. Accelerator of Antigen-Antibody Reactions

8. Stripping Reagent

9. Coloring Reagents

10. Chemiluminescence Reagents

D.

Mass Spectrometry

1. Reagents for Sample Pretreatment

2. Matrices for MALDI-TOF MS

3. In-gel Digestion Enzymes - Lysyl Endopeptidase -

4. LC/MS Solvents

5. Related Reagents

6. Peptide Calibration Standards of MALDI-MS

print (3.1 MB)





A.

Phos-tag™ - Phosphorylated Protein Analysis

39141.pngPhos-tag™is...

Phos-tag™ is a functional molecule that binds specifically phosphorylated ions.

Features

Selectivity of binding of a phosphate ion (2-) is much higher than that of other anions.

A stable complex is formed under physiological conditions (pH 5 to 8*).

*: In case of Zn2+ is used as M2+.

39155.png 

Description

Application

Phos-tag™ Acrylamide

Specific separation of phosphorylated proteins corresponding to the degree of phosphorylation

Phos-tag™ Biotin

Specific detection without any anti-phosphorylated antibodies on western blot

Phos-tag™ Agarose

Enrichment, separation and purification of phorphorylated proteins using column chromatography

Phos-tag™ Mass Analytical Kit

Analysis: Applicable to MALDI-TOF/Mass analysis of phosphorylated compounds in positive mode using an isotopic Zn2+-Phos-tag™ complex

Phos-tag™ was developed by Department of Functional Molecular Science at Hiroshima University.

1. Phos-tag™ Acrylamide (Please see the product list on the page #5.)

Separation of phosphorylated protein isoforms using SDS-PAGE

Phos-tag™ Acrylamide is added when a resolving gel of SDS-PAGE is prepared. Since Phos-tag™ in the gel can trap phosphorylated proteins and the migration speed of phosphorylated proteins decreases, they can be separated from non-phosphorylated proteins. (Phos-tag™ SDS-PAGE).

Features

Recognition of all phosphorylated forms of Ser / Thr / Tyr

Applicable to Western blotting and Mass analysis

Simultaneous detection of phosphorylated- and non-phosphorylated proteins using the total antibody without the anti phosphorylated protein.

39170.png 

Application

Time course of phosphorylation by using the Tyrosine Kinase Abl

Phosphorylated tyrosine was prepared by GST binding protein of tyrosine kinase Abl and the substrate peptide (Abltide)and separated with conventional SDS-PAGE and Phos-tag™ SDS-PAGE, respectively.

39409.png 

2. SuperSep Phos-tag™ - Precast Phos-tag™ Acrylamide Gels

This is a precast gel added with Phos-tag™ Acrylamide in advance. It can be used immediately after opening the package. It contains zinc as a supplemental metal. It has excellent storage stability by its neutral gel buffer. Sharp bands can be obtained.

39891.png 

Features

Phosphorylated- and non-phosphorylated proteins are efficiently separable with sharp bands.

Long-term stability (Stable for 6 months)

Physical Properties

Plate size

100 × 100 × 3 (mm)

Gel size

90 × 85 × 1 (mm)

Well number

13

17

Phos-tag™ conc.

50 μmol/L

Acrylamide conc.

10%, 12.5%, 15% or 17.5%

ZnCl2 conc.

100 μmol/L

Well volume

30 μL

25 μL

39949.png 

39968.png 

Description

Application

Wako Cat. No.

Package Size

SuperSep Phos-tag™ (50 μmol/L), 10%, 13 well

Separation of phosphorylated and non-phosphorylated proteins by SDS-PAGE

193-16711

5 sheets

SuperSep Phos-tag™ (50 μmol/L), 10%, 17 well

190-16721

5 sheets

SuperSep Phos-tag™ (50 μmol/L), 12.5 %, 13 well

195-16391

5 sheets

SuperSep Phos-tag™ (50 μmol/L), 12.5 %, 17 well

193-16571

5 sheets

SuperSep Phos-tag™ (50 μmol/L), 15 %, 13 well

193-16691

5 sheets

SuperSep Phos-tag™ (50 μmol/L), 15 %, 17 well

196-16701

5 sheets

SuperSep Phos-tag™ (50 μmol/L), 17.5%, 13 well

197-16851

5 sheets

SuperSep Phos-tag™ (50 μmol/L), 17.5%, 17 well

194-16861

5 sheets

Related Product

Description

Wako Cat. No.

Package Size

EasySeparator™

058-07681

1 set

40036.png 

3. Phos-tag™ Biotin

Detection of phosphoproteins for Western blotting

This is biotin-bound Phos-tag™ used for detection of phosphoprotein by Western blotting.

40256.png 

Features

All phosphoproteins can be detected.

Procedures of experiment are similar to those in ordinary Western blotting.

It can be conveniently used even when target anti-phosphorylated Thr/ Ser antibody is not available !

 

BTL-104 and BTL-105 have linkers with different length, but their usage are similar.

BTL-104 is recommendable as the first choice because of its high solubility.

4. Phos-tag™ Mass Analytical Kit

Improves Detection sensitivity of MALDI-TOF MS

Before use, Phos-tag™ Mass Analytical Kit is mixed with samples for MALDI-TOF MS. Phosphorylated molecule-Phos-tag™ complex is detected in a positive mode, and phosphorylated molecule usually difficult to detect can be detected with improved sensitivity.

Features

Detection sensitivity of phosphorylated molecule is improved.

Kit Contents

Phos-tag MS-101L 5 mg × 1 vial

([C27H29N6O64Zn2]3+ MW : 581.4)

Phos-tag™ MS-101H 5 mg × 1 vial

([C27H29N6O68Zn2]3+ MW : 589.4)

Phos-tag™ MS-101N 10 mg × 1 vial

(C27H29N6OZn2]3+ MW : 584.3)

 

101L, 101H and 101N have different types of Zinc.

101N contains many isotopes because it uses natural Zinc that has complex mass spectrum. It is suited for evaluation of condition.

101L and 101H contain Zinc with mass numbers of 64 and 68, respectively. They are suited for detection of phosphate group.

5. Phos-tag™ Agarose

Purification of phosphorylated protein by affinity chromatography

Fill Phos-tag™ Agarose in a column for use. It can be used for separation, purification and concentration of phosphorylated proteins. Because it is free from a surfactant or reducing agents, phosphorylated protein can be obtained in a condition similar to in vivo one.

Features

Phosphorylated proteins can be purified in 1 hour.

The proteins can be trapped in physiological condition (pH 7.5).

No reducing agent or surfactant is necessary.

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Description
(Manufacturer)

Grade

Manufacturer's

product No.

Wako Cat. No.

Package Size

Phos-tag™ Acrylamide
(Nard)

Separation of phosphorylated / non-phosphorylated proteins by SDS-PAGE

AAL-107M

300-93523

2 mg

AAL-107

304-93521

10 mg

Phos-tag™ Acrylamide 5 mM Aqueous Soln.

(Nard)

AAL-107S1

304-93526

0.3 mL

Phos-tag™ Biotin BTL-104
(Nard)

They can be used in place of anti-phosphorylated antibody. Any amino acids can be detected.

BTL-104

301-93531

10 mg

Phos-tag™ Biotin BTL-105
(Nard)

BTL-105

308-93541

10 mg

Phos-tag™ Biotin BTL-111

1mM Aqueous Solution
(Nard)

BTL-111 offers higher sensitivity than BTL-104.

BTL-111S1

308-97201

0.1 mL

Phos-tag™ Mass Analytical Kit
(Nard)

MALDI-TOF MS

MS-101KIT

305-93551

1 set

Phos-tag™ Agarose
(Manac)

Purification of phosphorylated proteins

AG-501

302-93561

0.5 mL

AG-503

308-93563

3 mL

B.

Protein Electrophoresis

1. SuperSep - Precast Polyacrylamide Gels

SuperSep is a precast polyacrylamide gel for electrophoresis of proteins or nucleic acids. Because the gel does not contain SDS, it can be used for SDS-PAGE in SDS-containing buffer, and for Native-PAGE in buffers that do not contain SDS. 12 kinds of precast gels are available.

B_well.psd 

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Product Specification

Well

13

17

Well volume (μL)

30

25

Mass of Total Protein (μg)*

3.3 ~ 6.5

1.3 ~ 3.9

Plate Size

100 × 100 × 3 (mm)

Gel Size

90 × 85 × 1 (mm)

*Target amount of protein that can be separated efficiently with SuperSep gel

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Target of separation

Please select appropriate gel concentration, with reference to the following separation pattern.

B_target.psd 

Description

well

Package Size

Wako Cat. No.

Shelf life

SuperSep Ace, 6%

13

10 sheets

195-15171

6 months

SuperSep Ace, 7.5%

13

10 sheets

198-14941

9 months

17

10 sheets

191-14931

SuperSep Ace, 10%

13

10 sheets

195-14951

17

10 sheets

192-14961

SuperSep Ace, 12.5%

13

10 sheets

199-14971

17

10 sheets

196-14981

SuperSep Ace, 15%

13

10 sheets

193-14991

17

10 sheets

190-15001

SuperSep Ace, 5-12%

13

10 sheets

199-15191

6 months

17

10 sheets

192-15201

SuperSep Ace, 5-20%

13

10 sheets

197-15011

9 months

17

10 sheets

194-15021

SuperSep Ace, 10-20%

13

10 sheets

191-15031

17

10 sheets

198-15041

SuperSep Ace, 15-20% (Tricine Gel)

13

10 sheets

198-15301

6 months

SuperSep, 12.5%, 2D

2D

10 sheets

190-13301

9 months

SuperSep, 5-20%, 2D

2D

10 sheets

197-13291

SuperSep, 10-20%, 2D

2D

10 sheets

192-14721

*Shelf life after manufacturing date

2. EasySeparator™

This is an electrophoresis tank for SuperSep precast polyacrylamide gel. Attachment and removal of the gel are easy.

Turn the knob after electrophoresis, then running buffer filled inside in the tank easily falls down. So, the gel can be removed directly.

Features

Easy set-up of SuperSep by turning the knob.

Easy removal of SuperSep without disposing the inside running buffer after electrophoresis.

Electrophoresis can be performed with a minimum amount of running buffer (250 ~ 350 mL)

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Description

Wako Cat. No.

Package Size

EasySeparator™

058-07681

1 set

3. Reagents for Gel Preparation

Followings are reagents used to prepare polyacrylamide gel for protein electrophoresis. Various kinds of reagents such as powder and ready-to-use solution are available.

Description

Grade*

Wako Cat. No. (Package Size)

Acrylamide, 99.0+% (cGC)

for Electrophoresis

017-08012 (25 g); 019-08011 (100 g); 011-08015 (500 g)

30 w/v% Acrylamide Solution, 37.5 : 1

016-15915 (500 mL)

Separating Gel Buffer Solution ( × 4)

(0.4 w/v% SDS in 1.5mol/L Tris-HCl, pH 8.8)

192-11041 (250 mL)

Stacking Gel Buffer Solution ( × 4),

(0.4 w/v% SDS in 1.5mol/L Tris-HCl, pH 6.8)

199-11051 (250 mL)

N,N'-Methylenebis (acrylamide) -HG,

99.0+% (Titration)

132-08171 (1 g); 130-08172 (25 g); 138-08173 (100 g)

N,N'-Methylenebis (acrylamide) -HG,

99.0+% (Titration)

for Molecular Biology

132-15082 (25 g); 134-15081 (100 g)

N,N,N',N'-Tetramethylethylenediamine

(TEMED), 99.0+% (cGC)

for Electrophoresis

205-06313 (25 mL)

Ammonium Peroxodisulfate (APS),

99+ % (Titration)

012-08023 (10 g); 016-08021 (100 g)

10 w/v% Ammonium Peroxodisulfate Soln.

(10 w/v% APS)

019-15922 (25 mL)

Riboflavin, 98.0+ % (Absorptiometry)

181-00581 (1 g)

2-Amino-2-hydroxymethyl-1,3-propanediol

(Tris), 99.0+ % (Titration)

for Molecular Biology

019-20091 (100 g); 011-20095 (500 g); 015-20093 (1 kg)

2-Amino-2-hydroxymethyl-1,3-propanediol

(Tris), 99.0+ % (Titration)

for Biochemistry

202-07881 (100 g); 204-07885 (500 g); 206-07884 (2.5 kg)

2-Amino-2-hydroxymethyl-1,3-propanediol 999,

99.9+ % (Titration)

015-16384 (100 g); 013-16385 (500 g); 011-16381 (1 kg); 017-16383 (5 kg)

Sodium Dodecyl Sulfate (SDS),

95.0+ % (GC) ; 98.0+ % (Titration)

197-07142 (25 g); 199-07141 (100 g); 191-07145 (500 g)

Sodium Dodecyl Sulfate (SDS), 95.0+ %

for Molecular Biology

190-13982 (25 g); 192-13981 (100 g); 194-13985 (500 g)

10% SDS Solution

Nippon Gene

311-90271 (100 mL); 313-90275 (500 mL)

2-Amino-2-Hydroxymethyl-1,3-propanediol

Hydrochloride (Tris-HCl)

for Biochemistry

010-17451 (100 g); 012-17455 (500 g); 016-17453 (1 kg)

2-Amino-2-hydroxymethyl-1,3-propanediol

Hydrochloride (Tris-HCl)

for Molecular Biology

015-20995 (500 g)

Tris-HCl Buffer Powder (1 mol/L, pH 8.0)

for Genetic Research

203-15361 (1 L×4)

1M Tris-HCl (pH 7.0)

Nippon Gene

311-90411 (100 mL); 313-90415 (500 mL)

1M Tris-HCl (pH 7.5)

316-90221 (100 mL); 318-90225 (500 mL)

1M Tris-HCl (pH 8.0)

312-90061 (100 mL); 314-90065 (500 mL)

1M Tris-HCl (pH 8.5)

314-90401 (100 mL); 316-90405 (500 mL)

1M Tris-HCl (pH 8.8)

311-90391 (100 mL); 313-90395 (500 mL)

1M Tris-HCl (pH 9.0)

314-90381 (100 mL); 316-90385 (500 mL)

Glycine 99.0+% (Titration)

 

073-00732 (25 g); 075-00731 (100 g); 077-00735 (500 g)

Tricine

Dojindo

341-02842 (25 g); 347-02844 (100 g)

* : for Molecular Biology - A series of reagents with DNase and RNase activities checked.

4. Protein Size Markers

Recombinant protein markers bound to stains in advance.

It is useful for confirming progress of electrophoresis or transfer efficiency.

41386.png 

No.

Description

Grade

Wako Cat. No.

Package Size

1

WIDE-VIEW™ Prestained Protein Size Marker III (11 ~ 245 kDa)

for Electrophoresis

230-02461

500 μL (for 100 tests)

2

WIDE-VIEW™ Prestained Protein Size Marker (15 ~ 140 kDa)

230-02221

500 μL (for 50 ~ 100 tests)

5. Premixed Buffers · Reducing Agents

This is a series of ready-to-use premixed buffers for protein electrophoresis. They can be used immediately after dilution.

Running Buffers for SDS-PAGE

Description

Application

Wako Cat. No.

Package Size

Composition

Running Buffer Solution ( × 10) (Tris / Glycine / SDS Buffer)

Laemmli method Tris-Glycine Running Buffer containing SDS

184-01291

1 L

Composition: 250 mmol/L Tris, 1,920 mmol/L Glycine, 1.0% (w/v) SDS

SDS-PAGE 10 × Running Buffer <Nippon Gene>

Laemmli method Tris-Glycine Running Buffer containing SDS

312-90321

1 L

Composition: 250 mmol/L Tris, 1,920 mmol/L Glycine, 1.0% (w/v) SDS

318-90323

5 L

10 × Tris-Glycine Buffer

Native-PAGE, Transfer Buffer

201-18601

1 L

Composition: 0.25 mol/L Tris 1.92 mol/L Glycin

Tricine Running Buffer Solution ( × 10) (Tris / Tricine / SDS Buffer)

Running Buffer for Tris-Tricine gel

200-17071

1 L

Composition: 0.5 M Tris / 0.5 M Tricine /1% SDS

Electrophoresis Sample Buffers

Description

Application

Wako Cat. No.

Package Size

Composition

Sample Buffer Solution (2-ME+) ( × 2)

Laemmli Sample Buffer with 2-Mercaptoethanol

196-11022

25 mL

Composition: 0.125 mol/L Tris-HCl (pH 6.8), 4w/v% SDS, 20 w/v% Glycerol,

0.002 w/v% BPB, 10 vol% 2-Mercaptoethanol

Sample Buffer Solution (2-ME+) ( × 4)

191-13272

25 mL

Composition: 0.25 mol/L Tris-HCl (pH 6.8), 8 w/v% SDS, 40 w/v% Glycerol,

0.02 w/v% BPB, 20 vol% 2-Mercaptoethanol

Sample Buffer Solution (2-ME-) ( × 2)

Laemmli Sample Buffer without 2-Mercaptoethanol

193-11032

25 mL

Composition: 0.125 mol/L Tris-HCl (pH 6.8), 4w/v% SDS, 20 w/v% Glycerol,

0.002 w/v% BPB

Sample Buffer Solution (2-ME-) ( × 4)

198-13282

25 mL

Composition: 0.25 mol/L Tris-HCl (pH 6.8), 8 w/v% SDS, 40 w/v% Glycerol,

0.02 w/v% BPB

Sample Buffer Solution with 3-Mercapto-1,2-propandiol ( × 2)

Laemmli Sample Buffer containing 3-Mercapto-1,2-propanediol (non-hazardous chemical) as substitute for 2-ME

199-16132

25 mL

Composition: 0.125 mol/L Tris-HCl (pH 6.8), 4 w/v% SDS, 20 w/v% Glycerol,

0.01 w/v% BPB, 10 v/v% 3-mercapto-1,2-propandiol

Sample Buffer Solution with 3-Mercapto-1,2-propanediol ( × 4)

196-16142

25 mL

Composition: 0.25 mol/L Tris-HCl (pH 6.8), 8 w/v% SDS, 40 w/v% Glycerol,

0.02 w/v% BPB, 20 v/v% 3-mercapto-1,2-propandiol

1× Buffer Series

The followings do not require dilution and are ready to use. The buffers are offered in sterile 5 L containers with capped stopcocks.

Description

Grade

Wako Cat. No.

Package Size

1×PBS(-)

for Biochemistry

164-25511

5 L

1×PBS(-)-T

161-25521

5 L

1×TBS

209-19121

5 L

1×TBS-T

206-19131

5 L

SDS-PAGE Buffer, pH 8.5

for Electrophoresis

192-16801

5 L

1×TAE

for Genetic Research

203-19141

5 L

Reducing Agents

This is a water-soluble reductant that cleaves disulfide bonds in proteins.

Description

Grade

Wako Cat. No.

Package Size

2-Mercaptoethanol

for Biochemistry

139-06861

2 mL×5A

137-06862

25 g

133-06864

100 g

(+/-)-Dithiothreitol

SH Groups Protection

047-08973

1 g

045-08974

5 g

041-08976

10 g

049-08972

25 g

041-08971

100 mg

3-Mercapto-1,2-propanediol

98.0+% (Titration)

207-09232

25 mL

201-09235

500 mL

TCEP Hydrochloride

for Biochemistry

205-18981

1 g

201-18983

10 g

0.5 mol/L TCEP Solution, Neutral

209-19241

1 mL

205-19243

5 mL

6. Staining Reagents

a. Negative Gel Stain MS Kit

It is known that protein bands separated by SDS/polyacrylamide gel electrophoresis (SDS-PAGE) can be visualized as transparent bands against the background of milky white gel stained by negative gel stain containing a Zn/imidazol reagent. We have improved the method using a new imidazol derivative reagent, which allows a clear and stable image of protein bands on the gel as sensitive as that by silver staining, in as little as 10 minutes. The staining technique is useful to obtain the clear and sensitive resolution pattern of the gel before immunoblotting as well as to excise and purify the band of interest from the gel without significant deterioration of amino acid residues for the subsequent studies of protein such as sequencing and mass analysis of peptide.

 

Staining Principle

Staining uses insoluble opaque ZnIm2 generated by reaction of free Zn2+ and imidazole.

Protein and protein-SDS complex react with Zn2+. The moiety reacted with Zn2+ remains clear because insoluble ZnIm2 does not deposit on it, but the moiety other than protein becomes opaque because ZnIm2 deposits on it. This type of staining is called negative staining, because unlike other staining, background is stained and protein is not stained.

41737.png 

Features

High sensitive (3 ng) and Quick detection (10 minutes).

Applicable to the subsequent studies of protein such as sequencing and mass analysis of peptide.

Repeatable staining and destaining are acceptable.

After destaining, the gel is applicable to Western blotting.

Kit Contents

Staining Solution A 500 mL × 1 bottle
Staining Solution B 500 mL × 1 bottle
De-staining Solution 500 mL × 1 bottle

Description

Grade

Wako Cat. No.

Package Size

Negative Gel Stain MS Kit

for Electrophoresis

293-57701

20 tests

b. Silver Staining Kits

3 types of high sensitive Silver Staining Kits are available.

Principle

Silver ion selectively binds to sulfhydryl group (-SH), amino group (-NH2), hydroxyl group (-OH), or carboxyl group (-COOH), which consist of protein. Especially, reaction with sulfhydryl group (-SH) occurs preferentially. Silver ion forming complex with ammonium ion in the reagent binds to protein, via substitution of ammonium ion and sulfhydryl group in protein.

29568.png Silver Stain MS Kit

This kit is produced for mass spectrometric sequencing of proteins separated by polyacrylamide gel electrophoresis, based on the method described by Shevchenko, et al. Due to the omission of glutaraldehyde treatment, proteins are rarely modified chemically and therefore can be detected at a sub-nanogram level on the electrophoretic gel.

Kit Contents

Enhancing Stock Soln.

200 mL × 1

Stopper

200 mL × 1

Staining Stock Soln.

200 mL × 1

De-staining Soln. A

50 mL × 1

Developing Stock Soln.

100 mL × 1

De-staining Soln. B

50 mL × 1

Developing Powder

20 g × 1

 

 

Application

MALDI-TOF MS of rabbit phosphorylase The band was excised and treated with Lysyl Endopeptidase® (#125-02543). Following the in-gel digestion and preparation, the sample was analyzed on MALDI-TOF mass spectrometer. (These data were provided by Dr. Y. Wada at Osaka Medical Center, Japan.)

 

41757.png 

Description

Grade

Wako Cat. No.

Package Size

Silver Stain MS Kit

for Electrophoresis

299-58901

20 tests

5510.pngSilver Stain Kit Wako

Silver Staining Kit Wako has high sensitivity and reproducibility. Staining finishes in 100 minutes.

41988.png 

Kit Contents

Fixing Stock Soln. (Glutaraldehyde) 200 mL × 2

Enhancing Stock Soln (Dithiothreitol) 10 mL × 1

Staining Soln. A (Silver nitrate) 100 mL × 1

Staining Soln. B (Ammonia & Sodium Hydroxide) 100 mL × 1

Developing Stock Soln. 100 mL × 1 (Formaldehyde & Citric Acid)

Description

Grade

Wako Cat. No.

Package Size

Silver Stain Kit Wako

for Electrophoresis

299-13841

for 10 sheets

5513.pngSilver Stain II Kit Wako

An improved type Silver Staining kit which can stain protein bands in 1 hour. It contains Stopper, which can be used for adjustment of staining for your intended use.

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Kit Contents (100 mL each × 1 bottle)

Fixing Stock Soln. (Methanol & Acetic Acid)

Enhancing Stock Soln. (Dithiothreitol & Glutaraldehyde)

Staining Soln. A (Silver Nitrate)

Staining Soln. B (Ammonia & Sodium Hydroxide)

Developing Stock Soln. (Formaldehyde & Citric Acid)

Stopper (Citric Acid)

Description

Grade

Wako Cat. No.

Package Size

Silver Stain II Kit Wako

for Electrophoresis

291-50301

for 10 sheets

42033.png 

Wako's Silver Staining Kits

Procedure

Reagent

Silver Stain Kit Wako supplies

Negative Gel Stain MS Kit

(Wako Cat. #293-57701)

Wako Cat. #299-13841

II

(Wako Cat. #291-50301)

MS

(Wako Cat. #299-58901)

Glutaraldehyde

Containing

Containing

Free

Free

Gel after electrophosing with SDS-Polyacrylamide gel

or Polyacrylamide gel

 

 

 

 

 

 

 

 

1. Fixing-1

Fixing Soln.-1

15 min. (Note 1)

10 min. (Note 1)

20 min. (Note 1)

Skip (Note 1)

 

 

2. Fixing-2

Fixing Soln.-2

15 min.

10 min.

10 min. (Note 1)

 

 

 

 

3. Wash

(Deionized Water)

5 min. × 3 times

 

10 min.

 

 

 

4. Enahncing

Enhancing Soln.

5 ~10 min.

10 min.

1 min.

 

 

 

5. Wash

(Deionized Water)

5 min.

5 min.

1 min. × 2 times

 

 

6. Silver Staining

Staining Soln.

15 min.

15 min.

20 min.

Staining Soln A

5~10 min.

 

7. Wash

(Deionized Water)

3~5 min. × 3 times

2~5 min. × 3 times

1 min. × 2 times

Wash

5 ~10 sec. × 3 times

 

8. Development

Developing Soln.

5 min.

5 min.

3 ~10 min.

Staining Soln. B

10~60 sec.

 

 

9. Stopping

Stopper

2~3 min. (Note 1)

2~3 min.

1 min.

 

 

10. Wash

(Deionized water)

2 min. × 3 times

2 min. × 3 times

1 min. × 3 times

Wash 1 min. × 3 times

 

 

(abt. 90 min.)

(abt. 70 min.)

(abt. 70 min.)

(abt. 10 min.)

Separation of gel (separation of target protein)

 

(Note 2)

(Note 2)

11. Destaining

 

 

 

15 min.

5 min. (Note 3)

 

 

 

 

 

In-gel Digestion (Note 4)

 

 

 

 

 

 

 

 

 

 

MS analysis

 

 

 

 

 

Kit Contents

(Note 5)

Fixing Solution

Fixing Stock Soln.

200 mL × 2

Fixing Stock Soln.

100 mL × 1

Enhancing Soln.

Enhancing Stock Soln.

10 mL × 1

Enhancing Stock Soln.

100 mL × 1

Enhancing Stock Soln.

200 mL × 1

Staining Soln.

Staining Soln. A

100 mL × 1

Staining Soln. B

100 mL × 1

Staining Soln. A

100 mL × 1

Staining Soln. B

100 mL × 1

Staining Stock Soln.

200 mL × 1

Staining Soln. A

500 mL × 1

Staining Soln. B

500 mL × 1

Developing Soln.

Developing Stock Soln.

100 mL × 1

Developing Stock Soln.

100 mL × 1

Developing Stock Soln.

100 mL × 1

Developing Powder

20 g × 1

Stopper

Stopper

100 mL × 1

Stopper

200 mL × 1

De-staining Soln.

De-staining Soln. A

50 mL × 1

De-staining Soln. B

50 mL × 1

Destaining Soln.

500 mL × 1

(Note 1)Since these kits do not contain this reagent, please prepare it by yourself. When you use Negative Gel Stain MS Kit, it is not necessary to fix samples. However, higher sensitivity will be obtained with 50 % ethanol fixing for 5 minutes

(Note 2)Brown colored staining parts are excised in case of silver staining. On the other hand, black colored parts against a black paper are excised in case of negative gel staining.

(Note 3)When used in subsequent MS analysis, de-staining is unnecessary. When used is subsequent Western Blotting, the gel is de-stained.

(Note 4)Lysyl Endopeptidase, Mass Spectrometry Grade (Wako Cat. No. 125-05061; 20 μg × 5) / Trypsin, from Porcine Pancreas, Mass Spectrometry Grade (Wako Cat. #202-15951; 20 μg × 5) are available from Wako. (Please see the page #22.)

(Note 5)Each Silver Stain Kit does not contain deionized water, methanol and acetic acid. Please prepare by yourself.

(The above mentioned reagents are not necessary when you use Negative Gel Stain MS Kit.)

c. CBB R-250 Stain Kits

5505.pngQuick CBB

Quick CBB is an upgraded version of the conventional CBB stain using CBB R-250. After fixing treatment, proteins are stained by immersing the gel in Quick CBB solution. Protein bands can be detected in a short time after SDS-PAGE treatment because no destaining procedure with acetic acid is necessary.

43236.png 

Features

Detectable in 50 minutes.

Easy preparation. (Prepare Staining Soln. by mixing Soln. A and B at a ratio of 1 : 1)

10 minute detection with microwave. (Application 2)

Description

Kit Content

Grade

Wako Cat. No.

Package Size

Quick CBB

1. Staining Solution A (1 L × 1)

2. Staining Solution B (1 L × 1)

for Electrophoresis

299-50101

2 L

 

5508.pngQuick-CBB PLUS

This is a protein staining kit which is a further upgraded version of Quick-CBB. In comparison with conventional Quick-CBB, fixing procedure is not required and organic solvents such as methanol and acetic acid are not used. The mixing procedure is unnecessary as all the required solutions for staining are contained in a single bottle. There are other improvements, such as the removal of background coloration. As in conventional Quick-CBB, destaining procedure is not required.

Features

No fixing procedure and no organic solvents are necessary.

A single-bottled ready-to-use protein staining kit

Simple and quick staining without the coloration on background.

Description

Grade

Wako Cat. No.

Package Size

Quick-CBB PLUS

for Electrophoresis

174-00553

250 mL

178-00551

1 L

43361.png 

C.

Western Blotting

1. Membranes for Blotting Applications

36528.pngClearTrans™ Membrane Series

ClearTrans™ SP PVDF Membrane, Hydrophobic

This PVDF membrane can be used for fluorescent protein detection.

It is used for highly sensitive detection of proteins due to its strong affinity with proteins. It also provides for a high S/N ratio because of the low background noise levels. It has small pores with diameters of 0.2 μm that reduce the permeability of proteins with low molecular weight.

ClearTrans™ Nitrocellulose Membrane

This product is Nitrocellulose Membrane for transcription. It can be used for colony/plaque lifts as well as nucleic acid blots. [Dimensions] 30 cm × 3 m, [Pore size] 0.2 μm

ClearTrans™ Nitrocellulose Membrane

This non-charged membrane for blotting applications is made of nylon 6-6. It can be used for colony/plaque lifts as well as nucleic acid blots.

Description

Grade

Wako Cat. No.

Package Size

ClearTrans™ SP PVDF Membrane, Hydrophobic, 0.2 μm

for Blotting

033-22453

1 roll (26 cm × 3 m)

ClearTrans™ Nitrocellulose Membrane, 0.2 μm

032-22663

1 roll (30 cm × 3 m)

ClearTrans™ Nylon Membrane, 0.45 μm

039-22673

1 roll (30 cm × 3 m)

2. Blocking Reagents

Blocking reagents used to prevent non-specific absorption of antibody to the transfer membrane. Generally, Bovine Serum Alubumin (BSA) or Skim Milk are used as a blocking reagent. Skim Milk is a potent blocking reagent, but it is not suitable for phosphorylated antibodies because they contain a lot of phosphorylated proteins.

26685.pngSkim Blocker

Skim Blocker is a blocking reagent that prevents non-specific absorption to the membrane or ELISA plates. This is a ready-to-use reagent that can be used immediately. Blocking finishes by shaking for 30 minutes at room temperature.

Description

Grade

Wako Cat. No.

Package Size

Skim Blocker

for Blotting

195-16455

500 mL

Skim Milk Powder

190-12865

500 g

Gelatin, from Bovine Bone*

for Biochemistry

074-02761

100 g

076-02765

500 g

Albumin, from Bovine Serum, Globulin Free*

019-15101

10 g

015-15103

50 g

013-15104

100 g

*: Not available for sale in the US.

3. Wash Solutions

Wash solution is a buffer for washing membranes for Western blotting or ELISA plates. It can be easily prepared by diluting the following solution with distilled water.

Description

Grade

Wako Cat. No.

Package Size

Composition

PBS-T, pH7.4 (×10)

for Blotting

163-24361

1 L

Composition: 1,370 mmol/L NaCl, 81 mmol/L Na2HPO4, 27 mmol/L KCl, 15 mmol/L KH2PO4,

1 (w/v)% Tween 20

TBS-T, pH7.4 (×10)

207-18061

1 L

Composition: 1,370 mmol/L NaCl, 27 mmol/L KCl, 250 mmol/L Tris, 1 (w/v)% Tween 20

10×TBS (pH7.4)

Nippon Gene

317-90175

500 mL

Composition: 1,370 mmol/L NaCl, 26.8 mmol/L KCl, 250 mmol/L Tris

20×TBS (pH7.4)

317-90371

500 mL

Composition: 400 mM Tris-HCl (pH 7.4), 3 M NaCl

10×PBS (−) (pH7.4)

314-90185

500 mL

Composition: 1,370 mmol/L NaCl, 81 mmol/L Na2HPO4, 26.8 mmol/L KCl, 14.7 mmol/L KH2PO4

 

4. Labeled 2nd Antibodies

Various types of labeled secondary antibodies are available. Anti-mouse IgG, HRP labeled antibody, anti-rabbit IgG and HRP labeled antibodies are now available.

44507.png 

Description

Grade

Wako

Catalog No.

Package Size

Anti Mouse IgG (H+L), Rabbit, IgG Whole, Peroxidase Conjugated

for Immunochemistry

012-23641

300 μL

018-23643

1 mL

Anti Rabbit IgG (Fc), Mouse, Peroxidase Conjugated

010-23941

300 μL

016-23943

1 mL

Anti Mouse IgG (H+L), Goat, IgG Whole, Biotin Conjugated, affinity purified

010-14031

1 mg

Anti Rabbit IgG (H+L), Goat, IgG Whole, Biotin Conjugated, affinity purified

013-14021

1 mg

5. Loading Control Antibodies

The followings are a monoclonal antibody for β-actin. A protein expressed by many eukaryotic cells, β-actin is widely used as a loading control in Western blotting.

Description

Grade

Wako Cat. No.

Package Size

Anti β-Actin, Monoclonal Antibody

for Immunochemistry

013-24553

200 μg

011-24554

1 mg

017-24556

5 mg

Anti β-Actin, Monoclonal Antibody, Peroxidase Conjugated

017-24573

200 μL

6. Antibody Detection Reagent

44643.pngMulti Capture HRP

This is a reagent kit binding to Fc region of antibodies to detect antibodies. Chemiluminescence can be used for detection because the capture reagent is HRP-labeled. Mouse IgG1 may be difficult to detect. In that case, use the Enhancer Reagent (for mouse) provided in the kit.

44658.png 

Features

Almost all antibodies can be detected. (Mouse IgG1 and Goat IgG may be difficult to detect.)

Sensitivity is similar or higher than that existing secondary antibodies.

Low lot-to-lot variation

44703.png 

Kit Contents

 

10 tests* (Wako Cat. #291-71801)

40 tests* (Wako Cat. #297-71803)

(1) Capture Reagent

50 μL

200 μL

(2) 20 × Reaction Buffer

10 mL

40 mL

(3) Enhancer Reagent for Mouse

50 μL

200 μL

*The package size is based on the use when capture reagent is diluted to 1 : 2,000.

Description

Grade

Wako Cat. No.

Package Size

Multi Capture HRP

for Blotting

291-71801

10 tests

297-71803

40 tests

 

7. Accelerator of Antigen-Antibody Reactions

This product is an accelerator to optimize antigen-antibody reaction for Western blotting, dot blotting, enzyme-linked immunosorbent assay (ELISA). Immuno-enhancer aids in obtaining high signal-to-noise (S/N) ratio.

Features

Enhances Immunoassay signals, effectively

High S/N Ratio

Applicable to Ready-to-Use Antibody Diluent

Kit contents

 

for 2 assays*

for 10 assays*

for 40 assays*

Reagent A

10 mL

50 mL

200 mL

Reagent B

10 mL

50 mL

200 mL

*Each package size shows an assay number when 5 mL each is used per assay.

45553.png 

45606.png 

Description

Grade

Wako Cat. No.

Package Size

Immuno-enhancer

for Blotting

294-68601

2 assays

290-68603

10 assays

298-68604

40 assays

Immuno-enhancer Reagent A

091-05811

200 mL

Immuno-enhancer Reagent B

098-05821

200 mL

 

8. Stripping Reagent

Stripping Solution is used to remove primary and secondary antibodies from a Western Blot membrane. Stripping is useful when several proteins are investigated on a sheet of membrane.

Usage

Wash a membrane reacted with a labeled antibody using TBS-T or PBS-T, and shake the membrane with Stripping Solution for 10 min. at RT.

Application

Sample: Mixture of 1 and 2

1. Transthyretin Variant (L55P), Human, recombinant (Wako Cat. #203-17321)

2. Glutathione S-Transferase

Membrane: PVDF Membrane; Blocking: 0.5% BSA

Labeled Antibody: Anti 6x His, MAb (9C11), HRP conjugated (Wako Cat.#010-23181) 4 mL

Chemiluminescent reagent: ImmunoStar LD (Wako Cat. #296-69901) Exposure time: 1 second

Wash the membrane with TBS-T

Incubate the membrane on an orbital shaker with Stripping Solution for 10 minutes

Wash the membrane with TBS-T

Blocking: 0.5% BSA

Labeled Ab: Anti Glutathione S-transferase, MAb, Peroxidase Conjugated

Chemiluminescent reagent: ImmunoStar LD (#296-69901); Exposure time: 1 second

45921.png

Description

Grade

Wako Cat. No.

Package Size

Stripping Solution

for Blotting

193-16375

500 mL

9. Coloring Reagents

36668.pngTMB Solution (for Membrane)

TMB (3,3',5,5'-tetramethylbenzidine) reacts with peroxidase to generate a bluish purple precipitate. This enables one-step detection of HRP-tagged antigens on membranes after Western blots, without the need for any special equipment.

Description

Grade

Wako Cat. No.

Package Size

TMB Solution (for Membrane)

for Blotting

208-19434

1 L

200-19433

250 mL

7311.png POD Immunostain Set

This is a sensitive enzyme-immunostaining kit of horseradish peroxidase-

labeled immunoglobulins on nitrocellulose membrane. Phenol is oxidized by reaction of phenol and hydrogen peroxide in the coloring reagent with POD, and blue-purple diformazan is generated in proportion to POD activity, in the presence of NADH and NTB. Antigen is detected by this reaction.

Kit Contents

NTB solution 250 mL × 1 bottle

NADH for 20 mL × 12 bottles

Substrate solution 13 mL × 1 bottle

Description

Grade

Wako Cat. No.

Package Size

POD Immunostain Set

for POD Staining

299-18841

20 mL × 12

7346.pngBCIP/NBT Solution

BCIP (5-Bromo-4-chloro-3-indolyl-phosphate) and NBT (Nitroblue Tetrazolium) generate an insoluble, dark blue-purple colored product by alkaline phosphatase. This reaction can be used for detection by Western blotting. The solution is prepared as a substrate for coloring in advance at a concentration suitable for immunoblotting.

This product is inapplicable to a microwell-EIA and immunohistostaining.

Description

Grade

Wako Cat. No.

Package Size

BCIP / NBT Solution

for Biochemistry

022-16231

100 mL

38297.pngPonceau 3R

The product is used for staining serum proteins by cellulose acetate membrane electrophoresis.

Description

Grade

Wako Cat. No.

Package Size

Ponceau 3R

for Electrophoresis

166-11921

5 g

164-11922

25 g

Ponceau-3R Stain Solution

169-18915

500 mL

 

10. Chemiluminescence Reagents

ImmunoStar™ LD

High-sensitive chemiluminescent detection in a short time

ImmunoStar™ LD (Long Detection), which is designed for a simple and highly sensitive immunoblotting, utilizing detection by enhanced chemiluminescence using our unique luminol derivative L-012 as the substrate can be detect labeled HRP.

Signal of Western blotting can be detected with high sensitivity due to

combination with unique enhancer. Also, it has long duration of luminescence.

46117.png 

Principle

L-012 is a luminol derivative, and dianion generated from H2O2 shows chemiluminescence.

46128.png 

Features

High Sensitivity (10-14; femto gram level)

Long term duration of chemiluminescence (CHL) (24 hours)

Low background (a high ratio of S/N)

Easy preparation (Prepare Luminescence Working Solution by mixing Soln. A and B at a ratio of 1:1)

High-sensitive detection in a short time

A good result with low background can be obtained by exposure for several seconds to minutes.

46166.png 

References

1. Obana, N., et al. : Mol. Microbiol., 77, 1416 (2010)

2. Fukumoto, H., et al. : J. Neurosci., 30, 11157 (2010)

3. Hyakkoku, K., et al. : Neuroscience., 171, 258 (2010)

4. Obana, N., et al. : J. Bacteriol., 193, 4417 (2011)

5. Kurotani, R., et al. : J. Biol. Chem., 286, 19682 (2011)

ImmunoStar™ Zeta

Long-lasting stable luminescence (mid-femtogram: 10-14g)

ImmunoStar™ Zeta using L-012, a luminol derivative, as a substrate. It can keep background noise low and detect chemiluminescence with a high S/N ratio. It also shows stable luminescence signals for a long period of time.

 

Features

A reagent that exhibits long-lasting stable luminescence

Low background (high S/N ratio)

2 component (equal proportions) type

46362.png 

ImmunoStar™ Reagents

A colorimetric method and UV detections are available after luminescence detection (picogram: 10-12g)

The ImmunoStar™ Reagents is a luminescent reagent whose detection limits for protein are in the picogram range.

A color and UV detection are available after luminescence detection. For UV detection, irradiate UV light after leaving the luminescent membrane for one night.

Features

A color and UV detection are available after luminescence detection.

Low background (high S/N ratio)

3 component type

Description

Characteristics

Detection Limit

Kit components

Grade

Wako Cat. No.

Package Size

ImmunoStar™ LD*

Useful where high sensitivity is required, such as in detecting underexpressed proteins.

low femtogram

10-15 g

2 components

(equal proportions)

for Blotting

296-69901

200 cm2

292-69903

1,000 cm2

290-69904

2,000 cm2

ImmunoStar™ Zeta*

A reagent that exhibits long-lasting stable luminescence

mid femtogram

10-14 g

2 components

(equal proportions)

291-72401

200 cm2

297-72403

1,000 cm2

295-72404

2,000 cm2

ImmunoStar™ Reagents

A colorimetric method and UV detections are available after luminescence detection

picogram

10-12 g

3 components

295-55201

1,000 cm2

291-55203

5,000 cm2

*: Not available for sale in the US and Europe.

D.

Mass Spectrometry

47096.png 

1. Reagents for Sample Pretreatment

Proteome Preparation Reagent

This product easily removes mucopolysaccharides and nucleic acids from extracted samples of plants, animals and microorganisms. As a result, the viscosity is decreased and the figure of electrophoresis is improved.

47155.png 

47173.png 

Description

Grade

Wako Cat. No.

Package Size

Proteome Preparation Reagent

for Proteome Research

164-22691

100 μL

Hydrophobic Protein Preparation Reagent

This product is used to accelerate solubilization of hydrophobic proteins and to obtain two-dimensional electrophoretic spots of proteins that cannot be isolated by a conventional method. It is also used to reduce the effects of lipids and nucleic acids contaminated in soluble proteins and obtain clearer spots.

47247.png 

Description

Grade

Wako Cat. No.

Package Size

Hydrophobic Protein Preparation Reagent

for Proteome Research

087-08891

0.2 mL

083-08893

0.5 mL

2. Matrices for MALDI-TOF MS

High-purity Matrix for MALDI-TOF MS

CHCA, SA and DHB

47471.png 

Features

1. High purity matrix is mixed with a test sample and used for proteome analysis by mass spectrometer (MALDI-TOF MS).

2. High purity matrix gives good mass spec reading because of its high purity by recrystallization).

Effects of recrystallization of α-Cyano-4-hydroxycinnamic Acid (CHCA)

When comparing with MALDI-TOF MS data for commercial CHCA, it was found that recrystallized CHCA (Wako Cat. No. 037-19261) gives clearer mass spec readings with less background noise. (These data were provided by Dr. Wada Y. at Osaka Medical Center, Japan)

47488.png 

Product List

Description

Grade

Wako Cat. No.

Package Size

α-Cyano-4-hydroxycinnamic Acid [CHCA]

for Proteome Research

037-19261

50 mg × 5

Sinapic Acid [SA]

192-13361

50 mg × 5

2,5-Dihydroxybenzoic Acid [DHB]

044-29101

50 mg × 5

2,5-Dihydroxybenzoic Acid Lithium Salt

048-29481

50 mg

2,5-Dihydroxybenzoic Acid Sodium Salt

041-29471

50 mg

3. In-gel Digestion Enzymes

Lysyl Endopeptidase, MS Grade / Trypsin, from Porcine Pancreas, MS Grade

Among the most important techniques in proteome analyses is the in-gel digestion of protein spots / bands that have been resolved by electrophoresis using digestive enzymes, such as trypsin and lysyl endopoptidase. Proteins can be identified by mass spectrometry of the peptides produced by in-gel digestion, and further information regarding post-translational modifications can be obtained.

Lysyl Endopeptidase, Mass Spectrometry Grade is a lyophilized product that retains sufficient activity for in-gel digestion and packed in very small quantities for convenience purposes.

47704.png 

Features

1. High specificity and efficiency of protein digestion allow for easy database searches by peptide mass.

2. Improved cleavage at lysine residue and increase in the number of peptides are obtained by combination with trypsin.

3. Packed in very small quantities in accordance with common usage guidelines, so sufficient activity is retained for in-gel digestion.

 

Comparison of In-gel Digestion Using Trypsin (Tp), Lysyl Endopeptidase (Lep) and Lep Combined with Tp (Lep +Tp)

BSA band (100 ng) resolved by SDS-PAGE was in-gel digested with Tp, Lep and Lep +Tp and analyzed by MALDI-TOF MS. The figure shows the individual mass spectra. The evaluation of these peptidases is summarized in the table.

Table: Comparison of Tp, Lep and Lep +Tp

These results indicate there are very few missed cleavages obtained by Lep digestion. When Tp is used concomitantly with Lep, missed cleavages decrease and the number of identified peptides increase compared to when only Tp is used.

 

Tp

Lep

Lep +Tp

Cleavage site

C terminal of Arg and Lys

C terminal of Lys

C terminal of Arg and Lys

Missed cleavage (Rates of missed cleavage)*1

Many (8 %)

Very few (0 %)

Few (3 %)

No. of identified peptides

17

19

22

*1The value resulted from subtracting the coverage obtained when database searches were performed with Missed cleavage 0 from that obtained when performed with Missed cleavage 1. "Coverage" is the percentage of peptides obtained after in-gel digestion in the whole sequence.

47716.png 

Description

Grade

Wako Cat. No.

Package Size

Lysyl Endopeptidase, Mass Spectrometry Grade

for Proteome Research

125-05061

20 μg × 5

Trypsin, from Porcine Pancreas, Mass Spectrometry Grade*2

202-15951

20 μg × 5

*2 : Not available for sale in the US

 

Related Products

Description

Grade

Wako Cat. No.

Package Size

Endoproteinase Asp-N, Sequencing grade

for Sequencing Grade

056-05921

2 μg

Endoproteinase Glu-C, Sequencing grade

050-05941

50 μg

V8 Protease [Endoproteinase Glu-C]

for Biochemistry

164-13982

2 mg

References

1. Wada, Y., and Kadoya, M.: J. Mass Spectrom., 38, 117 (2003).

2. Shevchenko, A., Wilm, MM., Vorm, O., and Mann., M.: Anal. Chem., 68, 850 (1996).

4. LC/MS Solvents

Liquid chromatography - mass spectrometry (LC/MS) is widely used in various fields including biological, food, and environmental analyses. In particular, recent breakthroughs in the development and upgrades of device interfaces have led to the use of LC/MS in microanalyses of environmental pollutants and chemical metabolites, etc.

Following products are ideal LC/MS reagent to analyze trace components.

47781.png 

 

Description

Wako Cat. No.

Package Size

Features

Specification

Acetic Acid

014-20063

1 mL × 5 A

. Suitability test for LC/MS performed

. Reduced background noise

Assay (HPLC): 99.5+%

Absorbance (1 4,250 nm): max. 0.50

Absorbance (1 4,254 nm): max. 0.10

Fluorescence test: to pass test

Suitability for LC/MS analysis: to pass test

018-20061

50 mL

Acetonitrile

016-19854

100 mL

. Suitability test for LC/MS performed

. Guarantees noise level at m/z 50~2,000

. Use of aluminum caps

Reduced risks of slight amounts of contaminants from plastic caps

Assay (cGC): 99.8+%

Density (20°C): 0.780 ~ 0.783 g/mL

Fluorescence test: to pass test

Suitability for LC/MS analysis: to pass test

012-19851

1 L

018-19853

3 L

Formic Acid

(abt. 99%)

063-04533

1 mL × 5 A

. Suitability test for LC/MS performed

. Reduced background noise

Assay (HPLC): 99.5+%

Solubility in water: to pass test

Absorbance (1 4,254 nm): max.1.00

Fluorescence test: to pass test

Suitability for LC/MS analysis: to pass test

067-04531

50 mL

0.1 vol% Formic Acid-Acetonitrile

062-04721

1 L

. Suitability test for LC/MS analysis performed

. Ready-to-Use eluent

Absorbance (200-400 nm): to pass test

Fluorescence test: to pass test

Water: max. 0.05%

068-04723

3 L

Methanol

132-14524

100 mL

. Suitability test for LC/MS performed

. Guarantees noise level at m/z 50~2,000

. Use of aluminum caps

Reduced risks of slight amounts of contaminants from plastic caps

Assay (cGC): 99.7+%

Density (20°C): 0.789~0.792 g/mL

Fluorescence test: to pass test

Suitability for LC/MS analysis: to pass test

138-14521

1 L

134-14523

3 L

2-Propanol

168-25531

1 L

. Suitability test for LC/MS performed

99.7+% (cGC)

Density (20°C): 0.784~0.787 g/mL

Fluorescence test: to pass test

Suitability for LC/MS analysis: to pass test

164-25533

3 L

Ultrapure Water

214-01301

1 L

. Decreased total organic carbon levels

. Guarantees the absorbance and fluorescence tests

. Use of specially processed glass containers /

aluminum caps

Density (20°C): 0.997 ~ 0.999 g/mL

Refractive index nD20: 1.332 ~ 1.334

Absorbance (210~400 nm): max. 0.01

Fluorescence test: to pass test

Total organic carbon (TOC): max. 4 ppb

210-01303

3 L

Related Products

Description

Wako Cat. No.

Package Size

Features

Joint Type

Wakopak®

MS-5C18GT

001-00030

2.0 mmf × 50 mm

. The glass lined inside wall of the stainless column allows

maximum inactivation

. Excellent peak shapes and recovery rates in analyses of trace

components within biological samples

DuPont

2.0 mmf × 100 mm

2.0 mmf × 150 mm

5. Related Reagents

Description

Grade

Wako Cat. No.

Package Size

Acetone

for Proteomics

015-22151

100 mL

017-22155

500 mL

Ammonium Hydrogencarbonate

018-21742

25 g

012-21745

500 g

Fluorescamine

060-05121

100 mg

Hydrochloric Acid

081-08671

100 mL

083-08675

500 mL

Iodoacetamide

099-05591

5 g

097-05592

25 g

Iodoacetic Acid

090-05641

1 g

098-05642

25 g

Phthalaldehyde (o-Phthalaldehyde)

160-23531

1 g

168-23532

25 g

Potassium Hydroxide

168-23571

100 g

160-23575

500 g

Sodium Azide

190-14901

1 g

198-14902

25 g

Sodium Carbonate

190-14842

25 g

194-14845

500 g

Sodium Thiosulfate

196-15162

25 g

190-15165

500 g

N,N,N',N'-Tetramethylethylenediamine

208-17452

25 mL

200-17451

100 mL

Thiourea

202-17352

25 g

206-17355

500 g

 

6. Peptide Calibration Standards of MALDI-MS

For MALDI-MS Calibration Standard

Peptide standards for matrix-assisted laser desorption ionization (MALDI) mass spectrometry used extensively for analysis of biological polymers such as proteins are highly purified meeting the requirements of MALDI-MS. They are available in step sizes of 500 in molecular weight for selection of calibrants corresponding to your sample.

Features

1. High Purity

2. Appropriate calibrant for your sample

3. Cost-conscious 2 nmol package

48299.png 

Description

(M+H)+

Grade

Wako Cat. No.

Package Size

Angiotensin II (Human)

<MALDI-MS Calibrant>

j 1046.5423

for Proteome Research

019-22931

2 nmol × 1 bottle

015-22933

2 nmol × 5 bottles

[D-Arg1, D-Pro2, D-Trp7,9, Leu11]-Substance P

<MALDI-MS Calibrant>

k 1497.8483

017-22971

2 nmol × 1 bottle

013-22973

2 nmol × 5 bottles

Apamin

<MALDI-MS Calibrant>

l 2026.8944

016-22941

2 nmol × 1 bottle

012-22943

2 nmol × 5 bottles

PAMP (Rat)

<MALDI-MS Calibrant>

m 2477.3642

160-24251

2 nmol × 1 bottle

166-24253

2 nmol × 5 bottles

ACTH (Human, 1-24)

<MALDI-MS Calibrant>

n 2932.5885

014-22981

2 nmol × 1 bottle

010-22983

2 nmol × 5 bottles

Adrenomedullin (Human, 22-52)

<MALDI-MS Calibrant>

o 3574.877

013-22951

2 nmol × 1 bottle

019-22953

2 nmol × 5 bottles

Insulin (Human)

<MALDI-MS Calibrant>

p 5804.6455

090-05881

2 nmol × 1 bottle

096-05883

2 nmol × 5 bottles

Listed products are intended for laboratory research use only, and not to be used for drug, food or human use.

Please visit our online catalog to search for other products from Wako ; www.e-reagent.com

This brochure may contain products that cannot be exported to your country due to regulations.

Bulk quote requests for some products are welcomed. Please contact us.

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