Mouse on mouse IHC staining (HepPar1, Clone OCH1E5, DAKO)
I'd like to check for HCC metastases in the lung. Therefore, I'm trying to estalish HepPar1 staining on mouse liver and lung FFPE tissue sections. Probably because of the primary antibody being produced in mouse I get unspecific staining in the lung.
What are your experiences with mouse on mouse IHC stainings: Is there a chance to get rid of unspecific staining and can anybody recommend a mouse IgG blocking reagent or kit?
In my experience the use of mouse monoclonal Antinodies does not work on mouse tissue, and in my knowledge it does not exist a blocking system of mouse epitopes. I suggest to' try rabbit monoclonal Or policlonal amtibodies, also for afp in the case of hcc.
If you use for your experiments immunodeficient mice (e.g. rag-/-) you have always a complete absence of breakground.
Thank you for your comments.
@ Atif Ahmed: Did you already use a mouse blocking kit that you would recommend? I came across a mouse on mouse polymer IHC kit being sold by abcam (ab127055). As it is quite expensive I would be happy if I could profit from your experiences.
@ Renato Franco: To my knowledge, at present, HepPar1 is the most reliable marker to detect HCC cells. Unfortunately, this antibody is only available as a mouse monoclonal antibody. Did you already stain for AFP and which antibody do you recommend?
@ Marco Chilosi: Unfortunately, I can't just switch to another mouse strain as I'm using a transgenic HCC mouse model.
I di not know the type of your research, but if you want only to' recognize hcc metastasis, that Are afp positive, in mouse lung, that is afp negative, afp could be' afp policlonal antibody could be' useful. I cannot help you for the product because i am not at work
HI, @Nora Hipp. I am running into the same problem that your originally described with high background from Hep-Par1. Did you finally figure it out. Thanks.
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