Recognizes a cluster of proteins between 70-80kD, identified as tyrosinase. Occasionally a minor band at 55kD is also detected. This monoclonal antibody shows no cross-reaction with MAGE-1 and tyrosinase-related protein 1, TRP-1/gp75. Tyrosinase is a copper-containing metalloglycoprotein that catalyzes several steps in the melanin pigment biosynthetic pathway; the hydroxylation of tyrosine to L-3,4-dihydroxy-phenylalanine (dopa), and the subsequent oxidation of dopa to dopaquinone. Mutations of the tyrosinase gene occur in various forms of albinism. Tyrosinase is one of the targets for cytotoxic T-cell recognition in melanoma patients. Staining of melanomas with this MAb shows tyrosinase in melanotic as well as amelanotic variants. This monoclonal antibody is a useful marker for melanocytes and melanomas.
Applications
Suitable for use in FLISA, Flow Cytometry, Immunofluorescence, Western Blot, Immunoprecipitation and Immunohistochemistry. Other applications not tested.
Recommended Dilution
Flow Cytometry: 0.5-1ug/10e6 cells Immunofluorescence: 1-2ug/ml Western Blot: 0.5-1ug/ml Immunohistochemistry (Frozen & Formalin fixed paraffin embedded): 0.5-1ug/ml/30min @ RT Immunoprecipitation: 1-2ug/500ug protein lysate Optimal dilutions to be determined by the researcher.
Positive Control
SK-MEL-13, SK-MEL-19, SK-MEL-30, SK-MEL-37 cells or Melanoma.
Storage and Stability
May be stored at 4°C before opening. DO NOT FREEZE! Stable at 4°C as an undiluted liquid. Dilute only prior to immediate use. Stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer. APC is light sensitive.
Note: Applications are based on unconjugated antibody.
Immunogen
Recombinant protein corresponding to tyrosinase.
Form
Supplied as a liquid in PBS, pH 7.2. Labeled with Allophycocyanin (APC).
Purity
Purified by Protein G affinity chromatography.
Specificity
Recognizes human tyrosinase, 70-80kD. Does not cross-react with MAGE-1 and tyrosinase-related protein-1, TRP-1/gp75. Staining of melanomas showed tyrosinase in melanotic as well as amelanotic variants.