[Histonet] Re: Histonet Digest, Vol 9, Issue 27

Donna L Brooks dlbroo01 <@t> gwise.louisville.edu
Tue Aug 17 10:50:59 CDT 2004

Hello Dr. WL Huang,

In regards to the mouse monoclonal antibody for p-c-Jun:
Be sure to check if your secondary antibody is "rat adsorbed".  I
suspect that you may be experiencing a cross reactivity due to the
species (rat/mouse)  being so closely related.  I have also done quite a
bit of immunoflourescent and immunoperoxidase staining of rodent neural
tissue from spinal cord injured rodents, and had this very same problem.
  Also, I would suggest checking the Ig groups.  For example, IgG versus
IgA.  Did you use a protein blocking solution?  If not you may want to
try that as well. Be sure to watch the concentration of your antibodies
and blocking serums/protein solutions.  If the concentrations are too
high (say 1:250 [high] vs  1:1000[lower]) you will get a high
background.  I suggest doing a titration method to see which
concentrations give th

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