Vector Laboratories
TMB Substrate Kit
SK-4400
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This is an easy to use TMB substrate kit for a number of applications. We use this TMB for use in sandwich ELISAs. Provided your upstream experiments work then this substrate will provide bright clear results.
ELISA
Antibody ELISAs with an HRP-conjugated secondary antibody and murine cells
Mix 2 drops Buffer stock, 3 drops TMB Stock solution, and 2 drops Hydrogen Peroxide solution into 5mL of milliQ water. DO NOT use PBS or other substrates to make substrate. For ELISA, do not use the stabilizing solution that comes in the kit. Pipette 50-100ul into each ELISA well and read when desired results are achieved.
If you are using a high a concentration of HRP-conjugated secondary, then TMB can react within seconds.
Results can be read with inactive HRP (bright yellow) at A450 after stopping with 1N sulfuric acid at A600.
N/A
Once your other assay is optimized this substrate reacts well with bright signal.
Can react too quickly with high concentrations of HRP.
If your assay is in order then this product will work for you.