Are all your TMB substrates transparent and do they all turn blue after reacting with the HRP enzyme of an antibody?
No, we offer different TMB substrates for different applications.
We recommend the following TMB substrates for this particular application:
- eBioscience TMB Solution (1X) (Cat. No. 00-4201-56)
- (For higher sensitivity) 1-Step Ultra TMB-ELISA Substrate Solution (Cat. No. 34029)
What is the shelf-life of 1-Step Ultra TMB-ELISA Substrate Solution?
The 1-Step Ultra TMB-ELISA Substrate Solution is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended. Please see section 8.1 of our Terms & Conditions of Sale (https://www.thermofisher.com/content/dam/LifeTech/Documents/PDFs/Terms-and-Conditions-of-Sale.pdf) for more details.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
My lab doesn't have a fluorescent or chemiluminescent plate reader and I need the most sensitive substrate for Horseradish Peroxidase (HRP). Which of your colorimetric substrates should I use?
The 1-Step Ultra TMB-ELISA is our most sensitive substrate for HRP in plate-based assays. It can detect down to ~ 2 pg/well or 20 pg/mL.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
Can the 1-Step Ultra TMB-ELISA Substrate Solution be used as a precipitating chromogen for Western Blot, Immunohistochemistry and immunocytochemistry?
No. This product is formulated to generate a soluble, colored product that absorbs light at 450 nm. When used properly, there will be no precipitate, therefore, this substrate is only appropriate for plate-based assays, such as ELISA.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
Is the 1-Step Ultra TMB-ELISA Substrate Solution an appropriate substrate for kinetic assays?
No. Although the non-stopped substrate will turn a blue color in the presence of Horseradish Peroxidase and absorb light at 307 nm and 652 nm, color development proceeds very quickly. In the time that it takes to read the full plate, the absorbance of the first wells will have changed. It is better to stop the the color development in all wells at approximately the same time point with an acidic soluton, turning it yellow and then read the absorbance at 450 nm.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.