The secondary antibody non-specific staining blues
Hello Gail and Kathleen
There are still people out there who are not at this congress. What you
seem to see is immunoglobulin staining due to crossbinding of your
secondary. You can do 2 things. First you order some antibodies which are
not, or only minimally crossreacting with immunoglobulins from other
species than the one they should recognise (Jackson has a whole panel of
these). A second way to deal with it is to first stain all your
immunoglobulins in the section with a FITC or Cy2 labeled antibody. In a
second step you than perform your matrix protein staining with a TRITC or
Cy-3 labeled antibody. Your specific staining should then be red while the
crossbound immunoglobulins should be green, or, yellow.
hope this works,
Jan
-----------------------------------------------------
Jan Bauer, Ph.D
Div. of Neuroimmunology
Brain Research Institute
Spitalgasse 4
A-1090 Vienna
Austria
tel: +43-1-4277 62813
fax: +43-1-4277 9628
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