Milk for blocking - (Jan/02/2006 )

Could you please inform me what kind of milk that we can use for blocking instead of Tween 20. How about concentration?
I always use tween 20 for blocking.

regard

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hi,

I always use 5% milk powder in PBS

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I always use 5% BSA in TBS-T for blocking of membranes to be used to detect phospho-proteins, and 5 % Milk in TBS-T for normal protein detection. But u can always use 5% Milk for phosphoprotein blot also, since BSA is more costly.

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Hello Ali,

U can use 5% non fat mild powder instead of BSA and 0.1 % Tween20 overnight(4 degree) for blocking.Addition of Tween 20 helps better in blocking.Cheers.

Saikat

Japan

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Hello
5% skimmed powder milk

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Don't use milk for blocking when you want to look for phosphoproteins, the casein in milk contains many phosphorylations and is often used as a positive control for phosphoprotein analysis.

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Also, don't use milk for blocking if you are using a biotin detection system. Milk contains biotin leading to a very high background.

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good to know (it's my next western-blot). thanks a lot.

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*blows the smoke off the barrel of his pistol*

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if ur talking about blocking the membrane for westernblot it good to use 3% skimmedmilk with 1.5%BSA for 1hr or vernight @4degree with out shaking.try and see.all the best.

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