Recipe

Immunoblotting blocking buffer

PBS

BSA (Fraction V, 3% w/v; optional)

Nonfat dry milk (5% wt/vol; optional)

Tween 20 (0.2%; optional)

Merthiolate (0.01%)

Prepare a solution of PBS with one of the following blocking agents:

BSA (Fraction V, 3% w/v). Store at 4°C.

Nonfat dry milk (5% w/v)

Tween(0.2%)

Nonfat dry milk (5% w/v)+ Tween (0.2%)

If the blocking buffer is not made fresh before each use, include 0.01% Merthiolate as a preservative. Sodium azide should be avoided, because it blocks horseradish peroxidase enzyme activity.

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  1. doi:10.1101/pdb.rec10056 Cold Spring Harb Protoc 2006: pdb.rec10056-

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