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文献和实验1. Materials(1) Antibody 7E3, 2L sup grown in flasks, frozen and thawed overnight.(2) BioRad Affi-Gel Protein A MAPS II Buffers cat. #1530-6160 ($161.00)(3) 50 mM Tris pH7.8.(4) 40 g ammonium sulfate for every 100 ml Sup.(5) 4L 100 mM Tris/pH
suspension. 5) Label another sample tube for each fluorochrome-conjugated antibody you are using, and add 1 drop of AbC™ capture beads (Component A in the AbC™ Anti-Mouse Bead Kit) to each tube. 6) Add a pre-titrated
Protocol for anti-HA antibody Western Blotting
seconds, and pour it off: this method was worked out in the Yamamoto lab eons ago). 7) Incubate with secondary antibody (anti-mouse) in 5% BSA/PBS/Tween as you would ordinarily. I usually go for 2 hours @ room temp. 8) 12-15 quick washes again(in 1XPBS
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