ab49387 staining MiTF (brown) in normal human skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 3% serum for 30 minutes at room temperature; antigen retrieval was by heat mediation in a buffer pH 9. Samples were incubated with primary antibody (1/25 in PBS + 3% BSA) for 1 hour. An undiluted biotin-conjugated horse anti-mouse IgG polyclonal was used as the secondary antibody. Melanocytes stained violet.See Abreview
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human malignant melanoma labelling MiTF with ab49387 at 1/10 dilution.
Overlay histogram showing MALME 3M cells stained with ab49387 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab49387, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.