[Histonet] xylene grades

[Rene J Buesa]

The xylene "grade" (if such a thing exists!) is of a lesser concern than assuring a complete dewaxing of the section. You have to completely eliminate all the paraffin.
  Using distilled water to later immerse the sections in a buffer (that contains salts) is of no relevance, because you are going to "salinize" the sections any way.
  I think that your problem is with your Ab supplier, or the lot. CD117 is a "tricky" one.
  I stopped having problems when I switched to DAKO Mo Ab and HIER at pH6
  René J.

Merced Leiker <leiker <@t> buffalo.edu> wrote:
  Would the grade of xylenes used for deparaffinizing make a difference for 
immunostaining of certain (picky) epitopes? We are using Certified A.C.S. 
xylenes.

What about rinsing in tap water vs. distilled water during rehydration?

The reason I'm asking is that we are having issues staining for c-kit (KIT, 
CD117) on FFPE tissue. We've tried all different antigen retrievals, 
including no retrieval, and still aren't seeing any stain.

Any help would be greatly appreciated. Thanks!


Merced M Leiker
Research Technician II
354 BRB (Lee Lab) / 140 Farber Hall (mail)
School of Medicine and Biomedical Sciences
State University of New York at Buffalo
3435 Main St, Buffalo, NY 14214
Ph: (716) 829-6033
Fx: (716) 829-2725


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