Hi, I've been using the monoclonal mix of Clontech against GFP. It works
perfect with high specificity at 1:100 and I bet it will work even at 1:200
or more diluted. The thing is also to dilute your cell extract appropiately.
With high expressing cell lines I easily can load as little as 1-5% of the
extract.
Detection is done with the ECL kit from Amersham which is also super.
Blocking step is 5% BSA. Rest is just TBS-Tween.
Martin Offterdinger <martin.offterdinger at akh-wien.ac.at> wrote in message
news:88jlmr$6qbi$1 at www.univie.ac.at...
> Hi!
> Are there any experiences with Clontech s a-GFP polyclonal AB in Westerns;
> They say it is suitable for westerns, but a get a very large number of
bands
> that make it impossible to distinguish which is the right one. Does one
have
> to use special blocking conditions or an extremely high dilution to
achieve
> some specificity?
> Is there anyone who has successfully used it for Western (Clontech, Cat #
> 8367-1)
> Thank you in advance
>>
