Reply: Negative slide has specific staining

From:Gayle Callis

Make sure your secondary antibody is adsorbed to mouse, we also use F(ab')
fragment of IgG to reduce any fc receptor problems, and dilute your
secondary in your normal serum blocking buffer.  % goat in normal block?
You can even use pure goat serum block, drain it off, add primary to that.
 You didn't say if your buffers/diluents contain detergent, Tween 20, this
has helped clean up slides for us.  
>Hi everyone
>I have a strange problem here.
>I'm doing some IHC for protein carbonyls on mouse lungs exposed to cigarette
>I'm blocking with goat serum.
>My primary antibody is Rabbit Anti-DNP
>My negative control is a slide that gets rabbit IgG instead of the primary.
>My secondary in biotinylated Goat Anti-Rabbit.
>Okay, my problem is that the ONLY slide that shows any of what appears to be
>specific staining (type 2 cells) is the negative slide (IgG instead of
>I've rerun it with the same results, so it's not that I got the slides mixed
>Any ideas?
Gayle Callis
Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367
406 994-4303 (FAX)

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