Western-Blot: PBS/TBS

Jürgen knobloch at uni-duesseldorf.de
Wed Sep 13 10:09:55 EST 2000


Hello!

Does anybody know if it makes a difference for certain antibodys (for
protein-detection  on a western-blot) to use PBS instead of TBS as basic for
all the incubation- and washing-buffers?

The reason why I´m asking is the following one:
Usually I use PBS - and there are no problems with background or anything
else.
But now I´ve tested a new antibody (an anti-phospho-tyrosine-antibody) and
I´ve followed excactly the manufacture´s manuel - this means I´ve used TBS.
The result was a very high backround (detection was done by CDP-Star) - so
that it is impossible to document the weak signals.

Because of a lack of protein-sample I can repeat this experiment only once.
And now I would like to use PBS instead of TBS.

Does anybody know a reason why this could be dangerous?

Thanx a lot,

Jürgen











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