Selected article for: "ice cold lysis buffer and lysis buffer"
Title: Triggering through CD16 or phorbol esters enhances adhesion of NK cells to laminin via very late antigen 6
  • Document date: 1992_11_1
    • ID: 3ymo8zw0_12
    Snippet: 32P Radiolabeling. Cells were washed in phosphate-free MEM containing 30 mM Hepes and 1% dialyzed FCS and starved for 1 h at 37°C . Cells were resuspended at 50 x 106/ml, incubated with 2 mCi/ml [32 p]orthophosphate for 4 h at 37°C, and then stimulated with the desired agent for the indicated times. Stimulation was terminated by adding ice-cold wash buffer (0 .4 mM EDTA-Na 2, 10 mM Na4P207, 10 mM NaF, 0.1 mM Na3VO4 in PBS, pH 7.4), pelleting ce.....
    Document: 32P Radiolabeling. Cells were washed in phosphate-free MEM containing 30 mM Hepes and 1% dialyzed FCS and starved for 1 h at 37°C . Cells were resuspended at 50 x 106/ml, incubated with 2 mCi/ml [32 p]orthophosphate for 4 h at 37°C, and then stimulated with the desired agent for the indicated times. Stimulation was terminated by adding ice-cold wash buffer (0 .4 mM EDTA-Na 2, 10 mM Na4P207, 10 mM NaF, 0.1 mM Na3VO4 in PBS, pH 7.4), pelleting cells for 5 min at 500 g, and resuspending them in ice-cold lysis buffer containing 1% /3-octylglucoside, 1 mM CaCl2, 1 MM MgC12, 0.1% NaN3 , 1 mM PMSF, 10 U/ml aprotinin, 10 ltg/ml leupeptin, 10 mM NaF, 150 mM NaCl, and 10 mM iodoacetamide for 30 min at 4°C.

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