Blocking Membranes for Western Blotting

darkwng · #1 (**permalink**) 06-26-2006, 07:54 PM

what is the best blocking agent for membrane western blotting?

is Blotto (milk) the best or casein and what about BSA??!

do i need to try them all first?

TerAle · #2 (**permalink**) 07-19-2006, 07:42 PM

5% BSA works well.

Alexei.

marcin · #3 (**permalink**) 08-10-2006, 07:58 PM

In my experience 5% milk in 1x TBS works the best. It is important to dissolve milk completely, otherwise milk clumps will cause nasty speckles on the film. If blocking is performed overnigt at 4C it is recommended to add some sodium azide.

sergbio · #4 (**permalink**) 02-07-2007, 06:36 PM

3% BSA work good. Two hours at 37 or 4C overnight

rimapant · #5 (**permalink**) 12-01-2007, 04:11 PM

My experience saysTBS/Casein from Bio-rad #161-0782 is best ( One hour at RT) Recently I tried Milk and BSA and casein all.

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Blocking Membranes for Western Blotting

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