[Histonet] Immunohistochemical detection of cytokines in older frozen sections

[Darius Alex Schneider]

Dear all,

I am trying to detect cytokines in human pancreatic sections from
diabetics. The blocks are at least 1y old, the sections are usually fresh.
I am fixing the slides with 2% PFA and permeabilizing with 0.1% saponin,
staining overnight in BSS-saponin with the primary, biotinylated antibody
and subsequently tyramid signal amplification and fluorescein based
visuatlisation. However, my results are very poor.
I am mainly interested in these cytokines:
IL1beta
IL1r
IL2
IL6
IL10
IL17
IL21
TNFalpha
IFNgamma

Is there somebody out there who is experienced in detecting these
cytokines, and could provide me with a comprehensive protocol? Also, who
knows what the best positive controls might be?

Thank you so much,

Darius
La Jolla, USA