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文献和实验Staining for Surface Receptors
coverslips. Equilibrate in incubator (~ 25'). 4. Add primary antibody at 1:500 to 1:1000 dilution to media for 30'. Example antibodies: M1 mouse anti-Flag antibody (Sigma , Covance ) (Ca+2 sensitive) 1:350-1:500; 12CA5 mouse anti-HA (Roche )1:1000
Flow Cytometry Assay to Measure Internalization of GPCR's
PBS Ca-free, Mg-free plus 0.04% EDTA to lift cells. 7) Pellet cells in centrifuge set at 4degC and rinse 1x with media. NOTE: Since the anti-FLAG M1 antibody ( Sigma , Covance )requires Ca ions to bind the FLAG epitope, you must rinse
Flow Cytometry Assay to Measure Internalization of GPCR
cells. 7) Pellet cells in centrifuge set at 4degC and rinse 1x with media. NOTE: Since the anti-FLAG M1 antibody ( Sigma , Covance )requires Ca ions to bind the FLAG epitope, you must rinse the pellet with media to get rid of any residual
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