Re: cytokine staining pattern in cells
|From:||Philip Oshel <firstname.lastname@example.org>|
Is there a full paper on this coming out? If so, or if it is out, I
would like a reprint, please.
>>Not interested in paraffin sections/antigen retrieved, only frozen
>>sections, and not formalin fixed, must be acetone or acetone/alcohol
>>fixation. Interested in IFN gamma and IL4
>>what kind of staining pattern are you seeing with cytokine staining?
>>Perinuclear staining? Many cells grouped in a pattern
>>Also, have you proven it is cytokine you have stained and not cross
>>reactivity of isotype matched control to cells within the tissue, eg
>>spleen? and did you do ligand block or a specificity control to prove it
>I am afraid I have a very sad message for you: forget about IFNg and IL4
>staining in acetone fixed tissue sections!
>We have just performed an extensive study using 13 different commercial
>available anti-IFNg antibodies. We obtained some hard evidence that it is
>not working: activated T-cells remain unstained and the positive signal you
>get is most likely nonspecific staining of a 5% plasma cell subset. We are
>very surprised that immunohistochemical staining of IFNg staining and other
>soluble cytokines have been applied by so many people and have published on
>it, sometimes even illustrated.
>A preface to our publication can be found in the Abstract book of the XIth
>Int. Congr. of Histochem. and Cytochem. last September in York, UK (pp.
>72-73, poster E20: Immunohistochemical visualization of IFNg: fake or
>fact?). An extensive publication is on its way to JHC.
>The only thing what is reliable in this respect is performing IFNg
>immunocytochemistry on intact cultured cells, preferably PFA-fixed and
>saponin in all your reagents.
>Best wishes, Chris
Supervisor, AMFSC and BBPIC microscopy facilities
Department of Animal Sciences
University of Wisconsin
1675 Observatory Drive
Madison, WI 53706 - 1284
voice: (608) 263-4162
fax: (608) 262-5157 (dept. fax)
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