[Histonet] Immunofluorescense staining

[John Kiernan]

If the antibody's supplier does not provide instructions for IHC you will need to test a wide range of concentrations of the primary antibody on sections known to contain the antigen in easily identifiable cell-types or other sites. Use a secondary antibody and detection system that you have already found reliable for primary antibodies raised in the same species as that in which the antibody for flow cytometry was raised. This would probably be a fluorescently labelled secondary antibody if "immunoflourescense" is specified.

The researcher probably needs to read a book about immunohistochemistry before expecting someone else to do it for her/him. An investment of $100 or so for a lab, along with a few hours of reading (by boss and workers), can save many hundreds of dollars that might otherwise be spent on antibodies and other reagents that don't work as expected. A good one to start with is Suvarna, S. K., Layton, C., Bancroft, J. D., eds. 2018. Bancroft's Theory and Practice of Histological Techniques 8th ed. London: Churchill Livingstone Elsevier. ISBN: 978-0-7020-6864-5. (NO, I don't have any vested interest!). I'm sure other histonetters will come up with similar and perhaps better suggestions.

Beware of working from "protocols" informally passed around among technicians, grad students and research fellows without published references (papers, books) that you can check before investing time, effort and money in a technique that's new to you. Informal lab notes are often locally treated as if they "have Authority,  not as the Scribes". In practical science it's probably best to go first with the Scribes, because their writings have been severely reviewed (if in good journals). Good books have references to peer-reviewed papers.

I could go on and on, but that's enough of my grumpy advice for now.

John Kiernan
London, Canada
https://www.schulich.uwo.ca/anatomy/people/faculty/emeriti/kiernan_john.html
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________________________________
From: Charles Riley via Histonet <histonet at lists.utsouthwestern.edu>
Sent: August 24, 2022 2:55 PM
To: histonet at lists.utsouthwestern.edu <histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Immunoflouresence staining

I have been given an antibody that is used for flow cytometry yet the
researcher wants to use it for IF staining.  The protocol they provided is
for the flow cytometry staining. Will this produce the immunoflourescense
results they are looking for or do i need to use a different method?
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