TMB Solution for Western Blotting
TMB (tetramethyl benzidine) solution is used with horseradish peroxidase (HRP)-based western blotting procedure, but not applicable for ELISA or immunohistochemistry.
Features
- Convenience: No CDD-camera-equipped imager or x-ray film needed
- Sensitivity: Low Picogram level
- Safety: Does not contain DAB or o-Dianisidine
- Ready to use: Does not need to dilute
Application: Detection for Gly m4, a major soybean allergen
| CBB Staining | Western Blotting |
|---|---|
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Experimental details
| Sample | Soy milk, dilution ratio as above |
| Detection | TMB Solution for Western Blotting; 5 minutes reaction time |
Data courtesy of Professor Tatsuya Moriyama and Dr. Erika Yano, Faculty of Agriculture, Department of Applied Biological Chemistry, Kinki University
Comparison with other picogram-level chemiluminescence kits and colorimetric substrates
| TMB Solution for Western Blotting; incubation time,10 minutes | |
|---|---|
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TMB Solution for Western Blotting |
| Other colorimetric substrates / incubation time,10 minutes | |
|---|---|
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Peroxidase Stain Kit（#26652-70） |
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Peroxidase Stain DAB Kit（#25985-50） |
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Peroxidase Stain DAB Kit + Metal Enhancer for DAB Stain（#07388-24） |
| Chemiluminescence kits / exposure time,10 minutes | |
|---|---|
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Chemi-Lumi One L（#07880） |
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Product A from competitor |
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Product B from competitor |
Experimental details
| Sample | Hela cell extraction, 2 times dilution started from 12 ug |
|---|---|
| Primary antibody | Anti-β-actin mAb manufactured by MBL, #177-3, X 5,000 dilution |
| Secondary antibody | Anti-mouse IgG, HRP-linked whole Ab Sheep manufactured by Cytiva, #NA931, X 10,000 dilution |
| Detector for chemiluminescence | LAS-3000 Super mode |
Quantitative analysis
Experimental details
| Sample | GST-His protein, 1.5 times dilution series started from 600 ng |
|---|---|
| Primary antibody | Anti-GST (mouse IgGa-k) Monoclonal(GS019) manufactured by Nacalai, #04435, X 2,000 dilution |
| Secondary antibody | Anti-mouse IgG, HRP-linked whole Ab Sheep manufactured by Cytiva, #NA931, X 10,000 dilution |
| Detection | TMB Solution for Western Blotting; 5 minutes incubation time |
| Analysis | Determination of band density with ImageJ software by Seiko Epson |
Application: Mid-femtogram-level sensitivity achieved with TMB Solution for Western Blotting along with Signal Enhancer HIKARI
| TMB Solution for Western Blotting; incubation time, 10 minutes | |
|---|---|
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TMB Solution for Western Blotting + Signal Enhancer HIKARI for Western Blotting and ELISA（#02270-81） |
| Chemiluminescence kits / exposure time, 10 minutes | |
|---|---|
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Chemi-Lumi One Super（#02230） |
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Product C from competitor |
Experimental details
| Sample | Hela cell extraction, 2 times dilution started from 3 ug |
|---|---|
| Primary antibody | Anti-GAPDH manufactured by Novous, #NB300-322, X 5,000 dilution |
| Secondary antibody | Anti-rabbit IgG, HRP-linked whole Ab Donkey manufactured by Cytiva, #NA934, X 10,000 dilution |
| Detector for chemiluminescence | LAS-3000 Super mode |
Video
TMB Solution for Western Blotting (5 minute incubation)
Download
TMB Solution for Western Blotting (Instruction)Ordering Information
| Product | Storage | Cat.No. | PKG Size | Price | |
|---|---|---|---|---|---|
| TMB Solution for Western Blotting | 4C | 18186-24 | 200 ml | 115.00 | Buy |