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Chemiluminescent reagents for western blotting ImmunoStar Series
ImmunoStar Basic
ImmunoStar Zeta
ImmunoStar LD

Immuno-Star series are chemiluminescent reagents for western blotting to detect the peroxidase (HRP, POD) labeling antibody.



1. Immuno-Star Series Selection Guid

Compared to our company's products.

Product Name ImmunoStar Basic ImmunoStar Zeta * ImmunoStar LD *
Type Cost, emission stability-oriented Balanced Emission intensity-oriented
Emission intensity (criterion of sensitivity) Medium sensitive
Capable of detecting the protein on the order of picograms (10-12).
High sensitive
Emission intensity of 4 to 20 times more than Basic.
Ultra-sensitive
Emission intensity of 20 to 100 times more than Zeta.
Emission stability
Cost
Membrane PVDF, nitrocellulose
Detection method CCD imager, X-ray film
Reprobing Possible
Features
  1. Emission stability is most excellent.
  2. Quantitativity is most excellent.
  3. Cost performance is excellent.
  1. Emission stability is excellent.
  2. Quantitativity is excellent.
  1. Series highest sensitivity.
Product Name ImmunoStar Basic ImmunoStar Zeta * ImmunoStar LD *
            *: Not available for sale in the US and Europe due to patents.
            Storage Condition: Keep at 2 ~ 10°C.

Immuno-Star series Criterion of emission intensity.



* Emission intensity represents the difference in signal intensity when it is detected under the same conditions.
The sensitivity of detection will change depending on the antibody concentration, antibody reaction time and exposure time, etc,.

2. Immuno-Star Basic

This is a luminescent reagent of medium sensitivity putting emphasis on cost performance, emission stability. For calm and stable emission persists, the emission signal can be easily adjusted by changing the exposure time. In addition, the calibration curve with a high linearity can be obtained in a wide range of the amount of protein.


Emission intensity and emission stability

Diluting FLAG-BAP gradually, emission intensity and emission stability were compared to other company's product A. Immuno-Star ® Basic showed a stronger emission signal than other company's product A (medium sensitive product), and also kept emission lasting stably.

Quantitative dynamic range

Diluting FLAG-BAP gradually, the quantitative dynamic range was compared to other company's product A. Immuno-Star ® Basic was able to create the calibration curve with a high linearity (more than R2=0.99) in a wider range of the amount of protein.

定量ダイナミックレンジ
Sample: FLAG-BAP
Gel: SuperSep Ace, 10-20%, 13well
    (Wako Catalog No.: 191-15031)
Membrane: ClearTrans SP PVDF Membrane, Hydrophobic, 0.2 μm (Wako Catalog No.: 033-22453)
Primary antibody: Anti DYKDDDDK tag, Monoclonal Antibody
  (Wako Cat. No.: 014-22383), 1,000-fold dilution
Secondary antibody: Anti Mouse IgG(H+L), Rabbit, IgG Whole, Peroxidase Conjugated (Wako Cat. No.: 018-23643), 10,000-fold dilution


* Please apply the quantitative analysis in the case of creating the standard curve to obtain a linear indicating the coefficient of determination of more than R2=0.99, and the target sample measured at the same time is within that range.
Product Name Package Size Wako Cat. No.
ImmunoStar Basic 200 cm2 (Luminescent liquid A: 10 mL, Luminescent liquid B: 10 mL) 295-75101
2,000 cm2 (Luminescent liquid A: 100 mL, Luminescent liquid B: 100 mL) 291-75103
5,000 cm2 (Luminescent liquid A: 250 mL, Luminescent liquid B: 250 mL) 299-75104
            Storage Condition: Keep at 2 ~ 10°C.

3. ImmunoStar Zeta

This is a high sensitive luminescent reagent that emphasizes each balance of emission intensity, emission stability, quantitative dynamic range.
While showing a strong emission signal, a stable emission persists.
In addition, the calibration curve with a high linearity can be obtained in a wide range of the amount of protein.


Emission intensity and emission stability

Diluting FLAG-BAP gradually, emission intensity and emission stability were compared to other company's product B(High sensitive product). Immuno-Star Basic showed an equivalent emission signal to other company's product B (High sensitive product), and kept emission lasting more stably."

Quantitative dynamic range

Diluting FLAG-BAP gradually, the quantitative dynamic range was compared to other company's product B. Immuno-Star Basic was able to create the calibration curve with a high linearity (more than R2 = 0.99) in the same range of the amount of protein as other company's product B.

Sample FLAG-BAP
Gel SuperSep (TM) Ace, 10-20 %, 13 well
  (Wako Cat. No. 191-15031)
Membrane ClearTrans® SP PVDF Membrane, Hydrophobic, 0.2 μm (Wako Cat. No. 033-22453)
Primary antibody Anti DYKDDDDK tag, Monoclonal Antibody(Wako Cat. No. 014-22383), 1,000-fold dilution
Secondary antibody Anti Mouse IgG(H+L), Rabbit, IgG Whole, Peroxidase Conjugated(Wako Cat. No. 018-23643), 10,000-fold dilution


* Please apply the quantitative analysis in the case of creating the standard curve to obtain a linear indicating the coefficient of determination of more than R2 = 0.99, and the target sample measured at the same time is within that range.

Comparison with the other company's products Emission intensity and emission stability

Emission intensity is the same level as other company's femtogram-level products (other company's product D, E). In addition, in the detection after 90 minutes from the reaction, it holds the emission intensity equivalent to other company's product D characterized by an emission durability.

Product Name Package Size Wako Cat. No.
ImmunoStar Zeta* 200 cm2 (Luminescent liquid A: 10 mL, Luminescent liquid B: 10 mL) 291-72401 *
1,000 cm2 (Luminescent liquid A: 50 mL, Luminescent liquid B: 50 mL) 297-72403 *
2,000 cm2 (Luminescent liquid A: 100 mL, Luminescent liquid B: 100 mL) 295-72404 *
            *: Not available for sale in the US and Europe due to patents.
            Storage Condition: Keep at 2 ~ 10°C.

4. ImmunoStar LD

This is a luminescent reagent of high sensitivity putting emphasis on emission intensity.
It is suitable for the detection of trace amounts of protein, such as lowly expressing protein.



Emission intensity



Sample FLAG-BAP
Gel SuperSep (TM) Ace, 10-20%, 17well
   (Wako Cat. No.: 198-15041),
Primary antibody Anti DYKDDDDK tag, Monoclonal Antibody
    (Wako Cat. No. 014-22383), 2,000-fold dilution
Secondary antibody Anti Mouse IgG (H+L), Rabbit, IgG Whole, Peroxidase Conjugated, 20,000-fold dilution(Wako Cat. No. 018-23643)
Exposure time 60 seconds (LAS4000, standard)

Immuno-Star LD was able to detect FLAG-BAP with a higher sensitively than other company's product B and ImmunoStar Zeta.

Emission stability



Diluting Anti-Mouse Immunoglobulins / HRP (DAKO, Code No.P0260) by 80,000-fold, and adding 1 μL of this to Fluorescens Luminescence Plate (Corning, Code No.3915), furthermore adding each 50 μL of luminescent reagents, then the emission stability was measured with the luminometer (TECAN Ultra Evolution).

Emission intensity after 120 minutes of the reaction was attenuated 61% compared to the emission intensity after 5 minutes of the reaction.

Quantitative dynamic range

Diluting FLAG-BAP gradually, the range of amount of protein, which can obtain the calibration curve with a high linearity, was examined. Immuno-Star LD indicated a high linearity(more than R2 = 0.99) in the range of amount of protein of 0.39 ~ 6.25 ng.
Since Immuno-StarLD has a very strong emission intensity and fast attenuation speed of signal, the quantitative dynamic range will be narrower than Immuno-Star Basic and Zeta.
Immuno-Star Basic and Zeta are recommended for quantitative analysis of the targeted antigen.

Sample FLAG-BAP
Gel SuperSep Ace, 10-20 %, 13 well (Wako Catalog No.: 191-15031)
Membrane ClearTrans SP PVDF Membrane, Hydrophobic, 0.2 μm (Wako Catalog No.: 033-22453)
Primary antibody Anti DYKDDDDK tag, Monoclonal Antibody(Wako Catalog No.: 014-22383), 1,000-fold dilution
Secondary antibody Anti Mouse IgG (H+L), Rabbit, IgG Whole, Peroxidase Conjugated
  (Wako Cat. No.: 018-23643), 40,000-fold dilution

Quantitative dynamic range
* Please apply the quantitative analysis in the case of creating the standard curve to obtain a linear indicating the coefficient of determination of more than R2=0.99, and the target sample measured at the same time is within that range.
Within the range of the amount of antigen 0.39 ~ 6.25 ng (Lane No.1 ~ 5), it showed more than R2 = 0.99 (1.2 order).

Product Name Package Size Wako Cat. No.
ImmunoStar LD* 200 cm2 (Luminescent liquid A: 10 mL, Luminescent liquid B: 10 mL) 296-69901 *
1,000 cm2 (Luminescent liquid A: 50 mL, Luminescent liquid B: 50 mL) 292-69903 *
2,000 cm2 (Luminescent liquid A: 100 mL, Luminescent liquid B: 100 mL) 290-69904 *
            *: Not available for sale in the US and Europe due to patents.
            Storage Condition: Keep at 2 ~ 10°C.

Immuno-Star Series Comparative Example

The followings show the difference of the emission intensity of Immuno-Star series and the change in emission intensity due to the exposure time. Please refer to them.

Sample FLAG-BAP
Gel SuperSep Ace, 5-20%, 17well (Wako Cat. No.: 194-15021)
Primary antibody Anti DYKDDDDK tag, Monoclonal Antibody (Wako Cat. No.: 014-22383)
Secondary antibody Anti Mouse IgG (H+L), Rabbit, IgG Whole, Peroxidase Conjugated (Wako Cat. No.: 018-23643)

5. Product List

Product Name Package Size Wako Cat. No.
ImmunoStar Basic 200 cm2 (Luminescent liquid A: 10 mL, Luminescent liquid B: 10 mL) 295-75101
2,000 cm2 (Luminescent liquid A: 100 mL, Luminescent liquid B: 100 mL) 291-75103
5,000 cm2 (Luminescent liquid A: 250 mL, Luminescent liquid B: 250 mL) 299-75104
ImmunoStar Zeta*
200 cm2 (Luminescent liquid A: 10 mL, Luminescent liquid B: 10 mL) 291-72401 *
1,000 cm2 (Luminescent liquid A: 50 mL, Luminescent liquid B: 50 mL) 297-72403 *
2,000 cm2 (Luminescent liquid A: 100 mL, Luminescent liquid B: 100 mL) 295-72404 *
ImmunoStar LD* 200 cm2 (Luminescent liquid A: 10 mL, Luminescent liquid B: 10 mL) 296-69901 *
1,000 cm2 (Luminescent liquid A: 50 mL, Luminescent liquid B: 50 mL) 292-69903 *
2,000 cm2 (Luminescent liquid A: 100 mL, Luminescent liquid B: 100 mL) 290-69904 *
            *: Not available for sale in the US and Europe due to patents.
            Storage Condition: Keep at 2 ~ 10°C.
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