[Histonet] von Willebrand factor IHC in rat kidneys
I'm dealing with a strange phenomena.
I have a nice working protocol for vWf in rat FFPE lung or heart, including a 15 min pretreatment of trypsin, 1 hour incubation of primary
antibody (Dako) 1:500
followed by Envision/HRP. I get constantly strong and clean staining.
The same protocol used for kidneys (yes, fixed and processed with the
exactly same schedule) shows only weak staining in the vessels and very
weak signal in the glomeruli.
I tried to use the primary more concentrated (up to 1:100) and to prolong
the its incubation to 2 hours with only little improvement.
Any ideas ? Should I consider another enzyme for pretreatment ? Longer
digestion with trypsin simply increased the background ,not the signal.
Is there any explanation why a protocol which works nicely in lung and
heart is not applicable for the kidneys?
Thanks to everybody,
Novartis Pharma AG
BU Transplantation Research
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