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Question about cell-based elisa protocol


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#1 SciCell

SciCell

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Posted 29 February 2012 - 04:23 PM

Hello all, I am coming back to this forum after a big gap. I normally assay my cels and clones for expression using FACS but as we are studying novel proteins, the commercially available antibodies are all not for FACS application. Many are for ELISA application. Can some one give me a protocol how to assay the cells for protein expression using elisa? assuming that the protein we are studying is expressed on or inside the cells and NOT a secreted protein.
I did ELISA many times before but for secreted proteins - I mean cell based (like the hybridomas) and even estimated the expression titer. But now I have the cells which express proteins ON the surface and I need to assay with a commercially available antibody using ELISA.
To start with - how do I coat the plates?

Thanks for your replies.




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