Protocol for F4/80 antigen

[mccullough_43]

I am fairly limited in my IHC experience and have a new antigen to stain with, F4/80 (rat anti mouse, serotec #mca497r).  The data sheet that came with the antigen suggests a protien digestion with trypsin.  I have not used this technique before and am hoping someone would have a protocol that I could follow to save me a lot of trial and error.  I am staining formalin fixed, parrafin embedded liver sections.

Thanks for any help or suggestions,
Sandi

[richard03]

The following is a protocol for your reference.
http://www.ihcworld.com/_protocols/antibody_protocols/f4.80_abd_serotec.htm

Hope this helps.

Richard

[ole]

Hi Sandi

There are many different enzymes you can use for antigen retrieval. They work in different ways but their ability to "retrieve antigens" are almost the same.  Not much difference between them for the majority of antigens where enzyme are the choice.

if you have other enzymes (eg Proteinase K RTU from DAKO) you should try them also. I have found Proteinase K to be better than trypsin (overall)., and its also easier to use as you dont have much experience yet.

Ole

[mccullough_43]

Thanks so much for the protocol and suggestions.  This is a great site!

[richard03]

Totally agree with Ole. Proteinase K is also my favorite enzyme and it can be used for almost all IHC application when enzyme digestion is needed. It is easy to make and often not over digest tissues.

Richard