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文献和实验Protocol for anti-HA antibody Western Blotting
room. Expose film for ~2 mins. Notes on the primary antibody: Source - Covance/Babco Description: Crude ascites fluid monoclonal anti-HA antibody (CATLN#135032002, 11mms-101R, 0.5 ml) Handling: Aliquoted into 10ul portions in ―20c
Western Blotting with Monoclonal Antibodies
% or 13% gels to PVDF membranes semi-dry transfer can also be used. Transfer proteins to PVDF membrane at 1.2 mAmp/cm2 for 1 hour and 45 minutes in transfer buffer (25mM Tris, 190mM glycine, 20% MeOH). Optional If blots are not to be used
Antibody Purification This protocol includes an ammonium sulfate cut, affigel blue chromatography and affinity chromatography. Solutions Affigel Blue Prewash 0.1 M acetic acid 5.7 ml glacial acetic acid
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