[Histonet] immunohistochemistry fish tissue - help
From: | "Tamara Franz-Odendaal" |
I have two questions:
1) Does anyone have a good protocol for immunohistochemistry working with fish tissue embedded in paraffin wax?
2) Any tricks with working with collagen antibodies (apart from hyaluronidase treatment)?
I have been trying without success to do the above, I have tried an Alexa 488 secondary as well as a horse radish peroxidase secondary. Neither are working and I am wondering if it is something peculiar to fish tissue. What are good blocks to use to prevent autoimmunofluorescence, or what secondary do you suggest. I am getting high backgrounds due to secondary. And also no signal in the area of interest (cartilage) so the primaries are also not working well. I am starting to wonder if the antibodies are at fault but before I order more wanted to know if anyone has any good working protocols.
I am working on larval tissue not embryonic. zebrafish.
Or do you suggest whole mount immunohistochemistry?
Thanks
Tamara
Dr. Tamara Franz-Odendaal
Dalhousie University
Halifax, Canada
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