Guest Mason Posted April 28, 2023 Report Share Posted April 28, 2023 Hello, I am having some issues with the HRP B cell ELISpot kit (3825-2H), where I did not get any distinct spots forming in the final step. I thought that since the Mabtech TMB it was not included in the kit, then any standard TMB would suffice? The two TMBs we have in our lab are the 1-Step TMB ELISA Substrate from ThermoFisher (Cat#34029) and the BD OptEIA TMB Substrate from BD (Cat#555214). What is the difference between these and the Mabtech ELISpot TMB? Attached is a picture of my ELISpot plate after addition of the BD OptEIA TMB Substrate. Some of the wells turned blue indicating the presence of antibody but I could not see distinct spots even after 30 mins. As our lab is based in New Zealand, it takes a long time to ship products from Mabtech so if there was an alternative it would be much appreciated! Thanks! Quote Link to comment Share on other sites More sharing options...
Christian@mabtech.com Posted April 28, 2023 Report Share Posted April 28, 2023 Dear Mason, welcome to the forum! Normal ELISA TMB substrate will not work in ELISpot. In order to generate spots you need a precipitating TMB substrate. Mabtech has such a substrate here: https://www.mabtech.com/products/elispot-substrate-tmb-hrp-3651-10-0 I am unfortunately unaware of alternatives, but reach out to our distributor in Australia! Quote Link to comment Share on other sites More sharing options...
Guest Mason Posted May 2, 2023 Report Share Posted May 2, 2023 Thanks. I see on the step-by-step guide to ELISpot on your website that AEC can be used as an alternative to TMB. Is there anywhere you can recommend purchasing this from if it's not a Mabtech product? Quote Link to comment Share on other sites More sharing options...
Christian@mabtech.com Posted May 2, 2023 Report Share Posted May 2, 2023 I have used BDs AEC ELISpot substrate and it works well with HRP detection mabs! https://www.bdbiosciences.com/en-eu/products/reagents/immunoassay-reagents/elisa/elisa-kits/aec-substrate-set.551951 Quote Link to comment Share on other sites More sharing options...
Guest Mason Posted May 2, 2023 Report Share Posted May 2, 2023 That's great, thanks for your help! Quote Link to comment Share on other sites More sharing options...
Christian@mabtech.com Posted May 4, 2023 Report Share Posted May 4, 2023 Excellent, any time! Quote Link to comment Share on other sites More sharing options...
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