Santa Cruz Biotechnology
Mast Cell Tryptase Antibody (AA1)
sc-59587
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Identification of mast cells in formalin-fixed paraffin-embedded human tissues is normally accomplished by specialized histological stains (toluidine blue, chloroacetate esterase) that bind to granule components. In this study, I wanted to assess the expression of other proteins in mast cells, and therefore needed an immunofluorescent approach to allow simultaneous analysis of multiple markers. Tryptase is a reliable marker by immunohistochemistry, and can be stained by the AA1 clone.
Immunofluorescence
FFPE human tissue (intestine, skin)
Epitope retrieval with citrate buffer, pH 6. Overnight staining at 4C with 1ug/ml primary antibody.
2% BSA with 0.1% triton X-100 in PBS.
Biotinylated rat anti-mouse IgG1, clone RMG-1, Biolegend, 2.5ug/ml for 1hr at room temperature.
Goat anti-rat IgG (H+L) Alexa Fluor 488, ThermoFisher, 10ug/ml for 30min at room temperature.
Nikon E800 Eclipse microscope
Bright annular staining with granules visible at high magnification. Modest background that could be from the RMG-1 anti-mouse IgG1 secondary (it isn't cross-adsorbed against human). Suitable for use in immunofluorescence. I tested epitope retrieval with citrate buffer pH 6 and EDTA pH 8. Both worked. Epitope retrieval may not be necessary for detection of tryptase, but it will be required for other antigens, and at least the tryptase epitope is robust. The staining also worked on frozen sections.
None
Very bright
Some background in the lamina propria of the intestine that could be improved with optimization.
Great antibody. Versatile.