The secondary antibody non-specific staining blues

From:Jan Bauer

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 Bauer, Ph.D
Div. of Neuroimmunology
Brain Research Institute
Spitalgasse 4
A-1090 Vienna

tel:    +43-1-4277 62813
fax:    +43-1-4277 9628

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