We are using a replication-competent virus to express an immune-stimulatory protein on cell surface. We want to determine the functionality of the virus-encoded protein. Is it ok to fix the virus-infected cancer cells with 4% PFA to inactivate virus and cells, and then use these fixed cells for stimulation of splenocytes? Could the PFA fixation effect effect structure of the protein and inhibit its function?
Hi Shyambabu,
No, the PFA will kill the splenocytes immediately (hoping you will use without dilution). And if you are using it at very high dilution, in that case also the PFA which acts as cross linker, can probably give you false positive readings.
I have used PFA in fixing my cells for IFA. In a time course experiment, whenever i fixed the cells in one column/row of 96-well plates and then cells (for next time-point) in other wells in the same plate behaved weird. We had to use chamber slides for time course studies, where we used one slide for one time course.
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