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DNA extraction for 8-OHdG assay
for DNA extraction from human & animal parenchyma tissue
DNA Extractor® TIS Kit
(Wako Catalog No. 296-67701;50 Reactions)
DNA Extractor® TIS Kit is specially designed for the extraction of DNA from human and animal parenchyma tissue. The
base of this kit is a procedure using Sodium Iodide (NaI) as a chaotropic agent, which allows for a safe extraction without
hazardous phenol/chloroform. The procedure using Sodium Iodide is an extraction procedure which causes less oxidation of
DNA. Additionally, the DNA Extractor® TIS Kit is able to inhibit the oxidation of DNA by using an Oxidation Inhibitor. It is
the most useful DNA extraction kit for the assay of 8-OHdG (8-Hydroxy-2'-deoxyguanosine), which is a marker of oxidative
stress.
Features
Useful for detection and determination of 8-OHdG, a marker of exidative stress.
Achieve inhibition of the further DNA oxidation by using an oxidation inhibitor
Suitable for a wide range of applications, which require DNA extraction from human and animal
parenchyma tissues.
DNA extraction from mouse liver and 8-OHdG Assay using the extracted DNA
DNA was extracted from a mouse liver using DNA Extractor® TIS Kit [Protocol 1] and DNA Extractor® WB Kit (Wako Cat. #291-50502).
Both of the obtained DNA were digested to mononucleotides with nuclease P1 using 8-OHdG Assay Preparation Reagent Set (Wako
Cat, #292-67801).
8-OHdG was determined by both of High Sensitive 8-OHdG Check (Wako Cat. #307-07921; ELISA method) and
HPLC/ECD and obtained values were compared.
Measurement by ELISA method
(using High Sensitive 8-OHdG Check)
Measurement by HPLC / ECD
DNA Extractor® TIS Kit
DNA Extractor® WB Kit
8-OHdG conc.
(nmol/L)
DG conc.
(µmol/L)
8-OHdG
/106dG
0.59
1.41
692
687
0.8
2.1
(HPLC condition)
Column: Wakopak® Wakosil-II 5C18 RS (ø4.6 mm x 150mm)
Eluent: 50 mmol/L Sodium Acetate (pH 5.2), 9% Methanol
Flow rate: 1 mL/min.
Absorbance: UV 260 nm
ECD Electorolytic Potential: +600 mV
It is known that 8-OHdG is generated by oxidation during the extraction
operation of DNA. However, DNA Extractor® TIS Kit could reduce the amount
of 8-OHdG generated during operation by more than half compared to the
DNA Extractor® WB Kit, by which the oxidation during operation is relatively
low. In addition, it was found that the concentration of 8-OHdG in
deoxyguanosine was very low. These results indicate that DNA Extractor® TIS
Kit has superior performance as a DNA extraction kit for 8-OHdG assay.
Kit Contents (50 Reactions)
Lysis Solution
Enzyme Reaction Solution
RNase Solution
Protein Digestion Solution
Oxidation Inhibitor
Sodium lodide Solution
Alcohol Solution
PEG Solution
2 bottles x 75 mL
1 bottle x 15 mL
1 vial x 50 µL
1 vial x 750 µL
1 vial x 350 µL
1 bottle x 15 mL
1 bottle x 30 mL
1 bottle x 20 mL
8-OHdG Assay Preparation Reagent Set
(Wako Catalog No.292-67801; 50 Reactions)
It is necessary to hydrolyze DNA extracted from tissues when
measuring 8-OHdG (8-hydroxy-2'-deoxyguanosine) in tissues.
The 8-OHdG Assay Preparation Set contains nuclease, buffer and
reagent which are used for the preparation of the assay of 8-OHdG
(8-hydroxy-2'-deoxyguanosine), a marker for oxidative stress. This kit is
able to reduce the variation of the enzyme reaction by addition of each
reagent to the sample DNA in accordance with the protocol. Therefore,
stable assay results for a marker of oxidative stress can be obtained.
Kit Contents (50 Reactions)
Acetic Acid Buffer
Nuclease P1
Tris Buffer
Alkaline Phosphatase Solution
1 vial x 950 µL
1 vial x 500 units
1 vial x 1 mL
1 vial x 50 µL
[Product List]
Description
Wako Cat. No.(Pkg. Size)
Application
DNA Extractor® TIS Kit
for Genetic Research; Keep at 2 ~ 10ºC
8-OHdG Assay Preparation Reagent Set
for Genetic Research; Keep at -20ºC
296-67701
(50 Reactions)
292-67801
(50 Reactions)
• DNA extraction from human/animal parenchyma tissue
• DNA extraction for 8-OHdG Assay
• Used for preparation of 8-OHdG assay
[Related Products]
Description
Wako Cat. No.(Pkg. Size)
Application
Nuclease P1, Penicillium citrinum
Keep at 2 ~ 10ºC; Lyophilized
Optimum temperature: 70ºC; Optimum pH: 5.3 (RNA and heat-denatured DNA)
High Sensitive 8-OHdG Check
Keep at 2 ~ 10ºC, manufactured by Nikken Seil Co.,Ltd.
Wakopak® Wakosil-II 5C18 RS (ø 4.6 mm x 150 mm) (D)
Wakopak® Wakosil-II 5C18 RS (ø 4.6 mm x 150 mm) (W)
145-08221
(500 units)
307-07921
(96 tests)
238-51471 (1 unit)
234-51473 (1 unit)
A kit content of DNA
Extractor® TIS Kit
ELISA kit for 8-OHdG
determination
HPLC Column
HPLC Column

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