[Histonet] CD4/CD8 staining in murine lungs
I am new to the world of Frozen section IHC and am trying to standardize a protocol for cd8/cd4 staaing in murine lung tissues embedded in OCT. I have tried without success to stain these sections with primary antibodies from biolegend, secondary antibody biotinylted anti-rat IgG(H+L) from vector labs. I was wondering if anybody has recomendations on a protocol or better antibodies than the one I am using. Also, in developing my slides with DAB (biogenex), sometimes my whole tissue sections turns brown within the matter of seconds. I have done through washes to remove any unbound antibody. I don't know why or where this is occuring. Is it in my snap freezing, sectioning, or staining? Thanks in advance.
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