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文献和实验Protocol for anti-HA antibody Western Blotting
) 5) Incubate in primary antibody solution: I use PBS with 0.1% TWEEN-20 plus 5% BSA as the basic solution, but you can use TBS too. I dilute the HA antibody 1:5000, just to be sure you will see a signal. I haven't EVER seen appreciable background. I've
Protocol for anti-HA antibody Western Blotting
it around for 5-10 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
. A rabbit polyclonal antisera is also available (101c500; BAbCO) but in our hands this was less reactive. The antibody cleanup protocol is now a seperate document Protocol Cells are grown in 5 mls of YPAD to an O.D. 600 of 0.75 to 1.
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