Can anyone guide me, What are all the things i've to concentrate more while preparing the samples for the detection of membrane bound proteins by indirect staining in flow cytometry?
Thank you for your suggestions Dominic.. Actually I'm going to check two different membrane bound protein expression on 1X10^6 cells/ml of human mast cell lines and I've purchased my protein specific primary antibodies, Isotype controls and their respective PE, FITC fluorescent secondary antibodies. Based on publication report they've used SIR buffer for resuspending the cell pellet and washing, staining procedures for 1 hr followed by fixing at 1 % paraformaldehyde. But here you have mentioned to avoid paraformaldehyde fixing, can u tell me the reason why we need to avoid fixing?
- Badges
- Science method