Improved technique utilizing nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose (original) (raw)

Gene Analysis Techniques, 1984

Abstract

A simple, economical, and efficient procedure for analysis of proteins (Western blotting) and DNA (Southern blotting) transferred to nitrocellulose for reaction with antibodies or nucleic acid probes is described. The techniques utilize nonfat dry milk as a protein-nucleic acid source for blocking nonspecific reactions, as an incubation medium, and for subsequent washing to remove unreacted reagents. The incubation cocktail, termed BLOTTO (Bovine Lacto Transfer Technique Optimizer), is superior to bovine serum albumin or gelatin for preventing nonspecific absorption in Western blot analyses and does not require the use of detergents or chaotropic agents to effect efficient reduction of background. BLOTTO, at the proper dilution in NaClNa citrate, is just as efficient in Southern blot analyses as more complicated cocktails typically used in the latter technique. We also found that BLOTTO works well for blocking, incubating, and washing ELISA plate assays relative to the normal BSA carrier, at a considerable savings to the laboratory.

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