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

  1. May 22, 2012 #1

    Dro

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    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.
     
  2. jcsd
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