Forums

GM

Guest Mason2023-04-28T01:52:11Z

---

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!



C

Christian@mabtech.com2023-04-28T08:05:02Z

Admin

---

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!

GM

Guest Mason2023-05-02T03:52:05Z

---

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?

C

Christian@mabtech.com2023-05-02T14:15:07Z

Admin

---

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

GM

Guest Mason2023-05-02T21:17:31Z

---

That's great, thanks for your help!

C

Christian@mabtech.com2023-05-04T13:42:08Z

Admin

---

Excellent, any time!