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How important is composition of TBST in diluting antibodies and Western Blotting?

 
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May22-12, 09:26 PM   #1
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How important is composition of TBST in diluting antibodies and Western Blotting?


I fear that I've added the wrong ingredient in making TBST and I'm not sure what the results will be like for my lab mates. Unfortunately I made 1M Tris-HCl instead of 10x TBS to add to the TBST. In other words, the pH is fine, but I have two times the amount of tris base added and none of the salt. (this is where it gets tricky) However 1L of my 1M Tris-HCl was added to 400mL of a previously fine 10x TBS. Of which I added 800mL to make 8L of TBST which was added to about 3L of stock TBST.

Basically, how badly did I mess up? Will it simply require more washing? Will it not work at all?
Any input would be greatly appreciated.

Thanks.
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blot, hcl, tbst, tris, western

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