I want to test siRNA or shRNA silence efficiency toward PD-1 in whatever cell lines, just to make sure that our siRNA sequence works. Which way is the easiest, WB, FACS or qRTPCR? which cell line should be used that produce much PD1? Can IFNγ stimulate T cell line to produce more PD1 which is just good to be used? Can I transduce virus to stimulate PD1 level, and then silence it? Thank you
Jennifer Gibbons
WB or qPCR both work- the ease depends on whether you have antibodies or primers to your protein of interest. ;) Knockdown by qPCR can be difficult to identify, as the Ct changes can be within variablility of the instrument (2-fold = 1Ct, the error range of the instrument). If you use qPCR, have a few replicates.
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