Protein will stripped by overwahsing? - (Oct/31/2007 )
QUOTE (Aimster @ Nov 1 2007, 03:47 PM)
Actually, long blocking times in milk CAN cause loss of antigen from the membrane - particularly if you are using a pretty high concentration of milk. It seems to be one of those quirky things about Westerns that few people are aware of. But if you've got plenty of antigen you're not likely to notice a problem. There's an old study that looked at this phenomenon specifically:
J Immunol Methods. 1989 Aug 15;122(1):129-35
http://www.ncbi.nlm.nih.gov/sites/entrez?D...Pubmed_RVDocSum
J Immunol Methods. 1989 Aug 15;122(1):129-35
http://www.ncbi.nlm.nih.gov/sites/entrez?D...Pubmed_RVDocSum
Interesting. I didn't know.
Thank you
-Missele-
Glad to be of service 
-Aimster-
Sometimes we have blocked overnight at 4ºC. I don't think it will strip the membrane
We have blocked with 1% milk without differences in background signal of higher percentage of milk. If the background is too high I try other reagent to block at same percentage, such as BSA.
-tonix37-