TMB Substrate Buffer is a single component, ready-to-use, working solution recommended for ELISA (microwell) procedures. TMB (3, 3′, 5, 5′-tetramethylbenzidine) is a commonly used substrate for the enzyme horseradish peroxidase (HRP) and the combination is one of the most acknowledged ELISA detection systems. Develops a soluble blue reaction product that may be read at 655nm. For endpoint assays the reaction may be stopped with acid, forming a yellow reaction product that may be read at 450nm or 405nm. Not recommended for membrane applications.
• This substrate is very light sensitive and must be protected from direct sunlight or any UV source.
• Bring to room temperature (approx. 25°C) before use. Note: Dilution of the substrate is not recommended. To reduce the intensity of a reaction, it is recommended that the antibodies or conjugates be diluted.
• Add 100 μl of TMB Substrate Buffer to each well and incubate for 5 –10 minutes at room temperature.
• A blue colored reaction product will develop, and negative control reactions remain clear non-colored solution in the wells.
• Add 100 μl of 1N H2SO4 to stop enzyme reaction after 10 minutes incubation. The blue color solution will turn to a yellow color.
•Blue reaction product, read at 655nm. Yellow, read at 450nm.
Store at 4° C
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.
Download SDS (MSDS)
Certificate of Analysis
Adobe Acrobat Reader is required to reliably view, print and comment on PDF documents
TMB Substrate Buffer, 1X
See how others have used TMB Substrate Buffer, 1X. Click on the entry to view the PubMed entry
bvseo-msg: The resource to the URL or file is currently unavailable.;
Rated 1 out of
Doesn't workTried it once. Even the standard curve of a commercial ELISA developed very little color. Decided to compare it to left over TMB solution of another ELISA kit. In the picture each of the 3 columns has the same standard curve. In both lateral columns I used the santa cruz TMB, in the middle one the left over TMB...
Date published: 2017-06-22
Rated 1 out of
Zhaomin Yang from
Poor qualityThe reagents were bad. We had to order them again from a different vendor for the experiment. This is awful.
Date published: 2017-02-08
Rated 5 out of
Various publications cite TMB Substrate BufferVarious publications cite TMB Substrate Buffer for its use in ELISA detection systems. Develops a soluble blue reaction product that may be read at 655nm. -SCBT Publication Review
To place an order using RMB or to ship to mainland China, please visit www.scbio.cn