Blocking buffer as antibody dilutent? - (Jun/09/2009 )
A Western Blot question. I use either fat-free milk or BSA as blocking (both in washingbuffer) and then this blocking buffer to dilute my antibodies, primary and second. Today I saw a protocol where they had milk as blocking but BSA in antibodydilutent? Is that something you all do? And in that case, why?
No, I do the same as you, milk as block and milk in the primary and secondary....
i use bsa and serum in my block and serum in my ab solutions (also, tween in ab solutions but not in block). but what you are using is just fine (if it works properly).
As long as it work, then it is OK to use blocking solution or BSA or even washing buffer. In my case, recently we bought secondary antibody that caused high background. By using blocking buffer (skim milk) it does reduced the background.
i do the same too. BSA and milk in blocking buffer and use it as dilutent for Abs.