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Western Blotting

32 bytes added, 00:54, 2 May 2009
→‎Protocol
==Protocol==
#Run SDS-PAGE gel and prepare transfer buffer
#Make sandwich, ensuring no bubbles between layers with black piece on bottom and layer as above. Place in apparatus so that the black sandwich touches the black transfer piece. Fill with transfer buffer. #Transfer 4h at 75V (in cold room) or overnight at 35V(on the bench).
#Carefully remove blot, stain with Ponceau solution and rinse with TTBS until all the red is washed off
#Block with 2% BSA in TBST or 5% skim milk powder in TBST for >1h