msd at ior.twmu.ac.jp (??) wrote in message news:<BD0761EB.894%msd at ior.twmu.ac.jp>...
> Hello.
>>> My problem is the 32kDa protein is supposed to be there,
>> and other experiments, RT-PCR, IHC, and DNA ladder assay
>> suggest the exhistance of the protein, but the objective band
>> in western blot , though it is supposed to be black on X-ray
>> film, is a white band just like a ghost in the night. Is there anyone
>> who has the right method to avoid this failure? I used goat IgG
>> for first antibody, and second antibody was Rabbit IgG anti
>> goat IgG. Please Help me!
Too much HRP, I believe, although some people say that too much
primary Ab can produce the same effect
see:-
http://www.piercenet.com/Objects/View.cfm?type=Page&ID=559F19D7-17FB-4596-BD0E-9123B5BEA481&Format=Print
HTH
David
