Looking for anticytokine antibodies

From:Gayle Callis

You can buy monoclonals from BD Pharmingen and affinity purified
polyclonals for RandD

Before you embark on IFNg staining, also IL4, you should read the
publication by Chris van der Loos et al, Immunohistochemical staining of
IFN gamma: Fact or Fake? J Histochemistry and Cytochemistry, 49(6):699-709,
2001.  I spent a week in his lab, repeating some of the experimental work
from this publication, cells stimulated to produce IFN gamma, make a pellet
in OCT, snap freeze and cryosection.  We did two fixations, acetone and
paraformaldehyde and saponin in all buffers and diluents. It was a
revealing story on poor to little staining of IFNg! You may have difficulty
getting any positive staining on frozen sections because of leakage of
cytokine from a cut cell, since the cytokine is located intracellularly at
the Golgi. 

His best results were with a polyclonal from R&D on cells pipetted onto a
to specially coated slides, and allowed to attach overnight.  We compared
these cells (never cut open) to frozen sections of cells expressing IFN
gamma.  The whole cells stained beautifully, frozen sections of cut cells
did not. This was similar to results we had in our lab, and frustrations
continue on how to solve the problem. 

Good luck! 

At 09:25 AM 9/10/02 -0400, you wrote:
>   Hi  all       CD161,  IL12p35, IL1p40, IL4, TNFa, IL10, IFN-g, TGFb     
> Just thought I would  ask....... TIA Luis 
Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

email: gcallis@montana.edu

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