Western blot analysis was performed on whole cell extracts (30 µg lysate) of A-375 (Lane 1), A-431 (Lane 2), B16-F10 (Lane 3), Caki-1 (Lane 4), Hep G2 (Lane 5) and COLO 205 (Lane 6). The blot was probed with Anti-Tyrosinase Mouse Monoclonal Antibody (Product # 35-6000, 2 µg/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjµgate (Product # A28177, 0.4 µg/ml, 1:2500 dilution). A ~80 kDa band corresponding to Tyrosinase was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product# SLF2000S). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Rat, Human, Not Applicable|
|Host / Isotype||Mouse / IgG2a, kappa|
|Immunogen||Recombinant tyrosinase protein.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (IHC)||Assay Dependent|
|Western Blot (WB)||2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Absence of recognition of common melanocytic antigens by T cells isolated from the cerebrospinal fluid of a Vogt-Koyanagi-Harada patient.
35-6000 was used in western blot to identify antigens recognized by autoreactive CD4 T lymphocytes isolated from a Vogt-Koyanagi-Harada patient who did not express HLA-DRB1*04:05
|Abad S,Wieërs G,Colau D,Wildmann C,Delair E,Dhote R,Brézin AP,Kawakami Y,Coulie PG,van der Bruggen P||Molecular vision (20:956)||2014|
Mitochondrial proteomics of the retinal pigment epithelium at progressive stages of age-related macular degeneration.
35-6000 was used in western blot to examine the subproteome of mitochondria isolated from human donor retinal pigment epithelium graded with the Minnesota Grading System
|Nordgaard CL,Karunadharma PP,Feng X,Olsen TW,Ferrington DA||Investigative ophthalmology and visual science (49:2848)||2008|
|Human||Not Cited||Immunophenotyping of melanomas for tyrosinase: implications for vaccine development.||Chen YT,Stockert E,Tsang S,Coplan KA,Old LJ||Proceedings of the National Academy of Sciences of the United States of America (92:8125)||1995|
|Human||Not Cited||Cultivation-dependent plasticity of melanoma phenotype.||Kodet O,Dvo¿ánková B,Krej¿í E,Szabo P,Dvo¿ák P,¿tork J,Krajsová I,Dundr P,Smetana K,Lacina L||Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine (34:3345)||2013|
|Human||Not Cited||T-cell immune function in tumor, skin, and peripheral blood of advanced stage melanoma patients: implications for immunotherapy.||Tjin EP,Konijnenberg D,Krebbers G,Mallo H,Drijfhout JW,Franken KL,van der Horst CM,Bos JD,Nieweg OE,Kroon BB,Haanen JB,Melief CJ,Vyth-Dreese FA,Luiten RM||Clinical cancer research : an official journal of the American Association for Cancer Research (17:5736)||2011|
||Immunophenotyping of melanomas for tyrosinase: implications for vaccine development.||Chen YT,Stockert E,Tsang S,Coplan KA,Old LJ||Proceedings of the National Academy of Sciences of the United States of America (92:8125)||1995|
|Flow cytometry assessment of residual melanoma cells in tumor-infiltrating lymphocyte cultures.||Richards JO,Treisman J,Garlie N,Hanson JP,Oaks MK||Cytometry. Part A : the journal of the International Society for Analytical Cytology (81:374)||2012|
||Flow cytometry assessment of residual melanoma cells in tumor-infiltrating lymphocyte cultures.||Richards JO,Treisman J,Garlie N,Hanson JP,Oaks MK||Cytometry. Part A : the journal of the International Society for Analytical Cytology (81:374)||2012|