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MICE ELISpot Assay

Guest Ivan Ssali

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Guest Ivan Ssali

Hi MabTech Respondent,

My name is Ivan Ssali from Medical Research Council (MRC/UVRI & LSHTM, Uganda).


We are working on a mice preclinical study and we recently bought a Mice ELISpot Kit (3321-2H).

It came along with a Datasheet & Protocol that we followed and used AEC substrate.

In the protocol it states that it was validated for mice spleen cells. We have also tried both fresh and cryopreserved cells but were unable to obtain spots in any of the wells, even the PHA well.

We therefore kindly request for a Zoom call in order to discuss the details so that you guide us on where we might have gone wrong.

Your timely response will be highly appreciated.




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Dear Ivan,


I highly recommend you purchase substrate from Mabtech. I also think it is best if you work with precoated kits.


We have not had any reported issues with our mouse antibodies so it is very unlikely a problem with the reagents.


Here are potential issues:

1. Something has gone wrong with the coating process. Do you employ etoh preactivation? What kind of elispot plates are you using?

2. The AEC substrate is not for ELISpot? You need to use precipitating substrate for spots to form. What supplier is the AEC substate from?


WE can have a zoom call no problem!





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