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Buffer solution used for making coating anti human IgG for B cells ELISpot

Guest Syabila2023-09-12T05:15:54Z

I would like to perform B Cells ELISpot using your product (Anti-human IgG mAbs (MT91/145), unconjugated. Would it be possible for you to explain how to prepare the coating buffer for making the coating solution? Is it okay to use the carbonate-bicarbonate buffer? It will be very helpful for us to the prepare the reagents. Thank you very much

Regards,

Syabila

admin2023-09-12T05:23:21Z

Admin

We recommend diluting the capture antibody in PBS. Concentration of the capture mab solution is generally 10-15ug/ml.

Add 100ul into each well of etoh activated PVDF plates.

Guest Syabila2023-09-12T06:28:55Z

Hi there

Just to make sure, do you mean PBS 1x? And I am also curious about what is the reason for using the PBS instead of carbonate bicarbonate buffer? Since we aware that PBS and carbonate-bicarbonate buffer have a very different pH where carbonate-bicarbonate buffer is more alkaline than PBS.

I also want to make sure, is the concentration of EtOH needed to activate the PVDF plate is following the protocol provided by the plate manufacture?

Thank you

Christian@mabtech.com2023-10-04T07:05:04Z

Admin

Yes I mean PBS 1x.

Antibodies are happier in PBS.

Yes, the etoh concentration needed for PVDF membrane activation is provided by Millipore. More and more people are switching to pre-coated ELISpot plates and this is my strong recommendation. Guarantees optimal results. ELISpot most often goes wrong due to incorrect coating effects.