white out

[David Cavanagh]

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