Fig. 2: Antibodies MCA-4H2 and RPCA-TH reliably detect both native and denatured TH in mouse and human tissue.
From: TNFα increases tyrosine hydroxylase expression in human monocytes
Human and murine brain sections (40 µm) were permeabilized, blocked, and stained with primary antibodies (MCA-4H2 and RPCA-TH) followed by HRP-conjugated secondaries and detected using diaminobenzidine enhanced with nickel (NiDAB, gray-black). a MCA-4H2 stains neuromelanin-expressing (brown) TH positive midbrain neurons and neuronal processes (gray-black) with no non-specific staining (secondary only, top panel; isotype control, second panel) in both human and murine tissues. b RPCA-TH shows similar highly specific staining of midbrain TH-positive neurons, confirming antibody specificity. a and b Human midbrain tissues showed as secondary-only and isotype controls exhibit endogenous neuromelanin (brown), not to be confused with immunostaining. c Western blot analyses of murine and human striatal tissues reveal similarly specific detection of TH (~63 kDa band) in both mouse and human, with minimal non-specific staining in negative control homogenate (parental CHO cell homogenate). (Left—MCA-4H2, right—RPCA-TH). d Blocking peptide/absorption control followed by western blot detection with either RPCA-TH and MCA-4H2 confirms the specificity of both antibodies for TH protein. HSP60 (loading control) is shown below and applies to c and d.
