[Histonet] PGP9.5 staining -- how to get rid of hair folliclebackground staining?

[Mitchell (Jean)]

A couple of questions for you first:  What is the thickness of your
sections? What concentration of primary do you normally use? 

I do PGP 9.5 staining on 50 micron frozen sections of skin. The hair
follicles will stain somewhat but should not be overwhelming.

Jean Mitchell
University of Wisconsin Hospital & Clinics
Department of Neurology
Madison, WI

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Cao,
Xudong
Sent: Friday, July 23, 2004 8:02 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] PGP9.5 staining -- how to get rid of hair
folliclebackground staining?

Dear all,
 
I am trying to stain for nerve fibers innervating the skin samples that
I have. While I am lucky to get the nerve fiber singnals, I am also
getting overwheelingly strong background staining from the hair
follicles. I know the follicle stains are background since they are also
stained in both of my controls where primary and secondary are omitted
are also stained. My question is: how to get rid of the hair follicle
stains? any suggestions? I am using Vector's ABC and SG substrate.
Thanks
 
Xudong Cao, Ph.D.
Biomedical Center
Brown University