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Blocking Membranes for Western Blotting

Blocking Membranes for Western Blotting - Western Blot Forum

Blocking Membranes for Western Blotting - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


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  #1  
Old 06-26-2006, 07:54 PM
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Question Blocking Membranes for Western Blotting



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?
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Old 07-19-2006, 07:42 PM
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Default Re: Blocking Membranes for Western Blotting

5% BSA works well.

Alexei.
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Old 08-10-2006, 07:58 PM
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Default Re: Blocking Membranes for Western Blotting

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.
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Old 02-07-2007, 07:36 PM
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Default Re: Blocking Membranes for Western Blotting

3% BSA work good. Two hours at 37 or 4C overnight
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Old 12-01-2007, 05:11 PM
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Default Re: Blocking Membranes for Western Blotting

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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Old 07-08-2009, 03:55 PM
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Default Re: Blocking Membranes for Western Blotting

I have tried all.
I would prefer TBS/Casein from Bio-Rad
Catalog # 161-0782

Good Luck
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