TMB is much more sensitive (factor 10). It's available as ready to use solution: http://www.seramun.com/products/substrates/horseradish-peroxidase.html
TMB is much more sensitive (factor 10). It's available as ready to use solution: http://www.seramun.com/products/substrates/horseradish-peroxidase.html
TMB (tetramethyl benzidine read at OD 450) has the advantage of being non carcinogenic and better when working with undergraduates. However, it can fall out of solution and usually requires gentle shaking during development. OPD (orthophenylene diamine read at OD 490) is the more traditional substrate. It does not fall out of solution, but is carcinogenic. Sigma also has a substrate solution that already contains the peroxide needed for color development.
Hi Bhagya, i totally agree with Diana explanation for recommendation of TMB
OPD and TMB. TMB is better.
Read TMB at 370 nm before addition of H2SO4. If you don't have 370 nm on your specrofotometer; use 405 nm. After H2SO4; 450 nm.
In compared to OPD, in our lab TMB gives more background. Has any one any expreience which company provide better TMB with less background?
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