Preparing blocking solution for Western Blotting - what milk power to use? - western blot (Sep/15/2006 )

i did my western blotting today, i used the blocking solution containing the nonfat powdered milk(i brought the "calcium plus non fat milk powder"), however, my supervisor told me that i should use the "calcium plus" and "nonfat" powder, he told me that i should buy the normal one which is not the nonfat and calcium plus, can anyone tell me that

1. why should use the nonfat (most journals pointed out) milk powder?

2. Did the "calcium plus" milk powder affect the blocking?

3. How do "fat" milk powder affect the blocking?

4. Would anyone can suggest me to buy which brands of milk powder are most suitable for blocking?

Thanks for helping,since i am so confused on this

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Hi!
I am new on WB, but what I can tell you is that I use nonfat powdered milk and bifidus free. I made it at 5% on TBS buffer and it works fine.
Good luck!

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if it contains the bifidus , does it affect the blocking?Thanks

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well, some colleagues recommend me that about bifidus, but I can´t explain exactly if it is very important for the blocking of if the reason is that it can contaminate the membrane. I will ask them and as soon as I can I will post it here.

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we use CARNATION non-fat milk powderfrom Nestle for blocking

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have calcium plus?

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no, just the regular one.

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Did the calcium rich powder affect the blocking?

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Fat does affect the blocking. One of my friend had bought the full cream milk powder and her western blot looked funny.

Hope this may help.

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I dont think we ever tried the calcium rich powder.

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