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  • Immunology
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  1. Hi All, I am just starting out with B Cell ELISpots, and just trying to get the assay running with the positive control (Total Ig) at this stage. (This is the first time B Cell ELISpots are being used in my lab, but we have successfully used T Cell ELISpots). I am using PBMCs from patient blood samples 6-9 days post infection. I have tried using the basic protocol provided by Mabtech. I have also tried stimulating with IL-2 and R848 for 48 hours. Finally I have also attempted substituting wash buffers with PBS, PBS/EDTA and PBS/Tween based buffers which I obtained from a different lab which had success with B Cell ELISpots. I am not seeing any spots. Any suggestions on what steps I should be looking at this point to troubleshoot? Thanks, Shawn
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