Key features and details
- Mouse monoclonal [HMB45 + M2-7C10 + M2-9E3 + T311] to Melanoma
- Suitable for: Flow Cyt, ICC/IF, IHC-P, IHC-Fr
- Reacts with: Human
- Isotype: IgG
Product nameAnti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3 + T311]
See all Melanoma primary antibodies
DescriptionMouse monoclonal [HMB45 + M2-7C10 + M2-9E3 + T311] to Melanoma
SpecificityThe HMB45 clone reacts with a neuraminidase-sensitive oligosaccharide side chain of a glycoconjugate present in immature melanosomes. The HMB45-reactive antigen is present in cutaneous melanocytes, prenatal and infantile retinal pigment epithelium and melanoma cells and is thought to be oncofetal in nature. This antibody has been shown to label the majority of melanomas. MART-1 recognizes a protein of 18kDa, identified at MART-1 (Melanoma Antigen Recognized by T cells 1) or Melan-A. Melan-A is a useful addition to melanoma panels as it is apparently specific for melanocytic lesions. Studies have also shown that MART-1 is more sensitive than HMB45 when labeling metastatic melanomas. Tyrosinase is a key enzyme involved in the initial stages of melanin biosynthesis. Studies have shown Tyrosinase to be a more sensitive marker when compared to HMB45 and MART-1. It has also shown to label a higher percentage of desmoplastic melanomas than HMB45.
Tested applicationsSuitable for: Flow Cyt, ICC/IF, IHC-P, IHC-Frmore details
Species reactivityReacts with: Human
HMB45 - Pigmented melanoma metastases from LN MART-1 - Recombinant human MART-1 protein Tyrosinase - Recombinant tyrosinase protein
- Metastatic melanoma in lymph node.
Please note that this antibody is an oligoclonal antibody. It is a cocktail of monoclonal antibodies that have been carefully selected. Oligoclonal antibodies have not only the specificity and batch-to-batch consistency of a monoclonal antibody, but also have the advantage of the sensitivity of a polyclonal antibody due to their ability to recognize multiple epitopes on an antigen.
The combination of HMB45, MART-1 (M2-7C10 + M2-9E3) and Tyrosinase (T311) (SwissProt: P14679 Human, Omim: 606933 Human, Entrez Gene: 22173 Mouse, Entrez Gene: 308800 Rat) make this quadruple antibody cocktail a first-order pan melanoma screener, and may prove to be a valuable marker for melanoma metastasis in sentinel lymph nodes (see reference 3.).
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.30
Constituents: PBS, Tissue culture supernatant
Proprietary preservative that is not sodium azide or thimerosal, protein carrier.
Concentration information loading...
Primary antibody notesThe combination of HMB45, MART-1 (DT101 + BC199) and Tyrosinase (T311) make this quadruple antibody cocktail a first-order pan melanoma screener, and may prove to be a valuable marker for melanoma metastasis in sentinel lymph nodes (see reference 3.).
Clone numberHMB45 + M2-7C10 + M2-9E3 + T311
Light chain typekappa
Our Abpromise guarantee covers the use of ab733 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|IHC-P||1/25 - 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
It is sometimes difficult to interpret DAB stained melanomas due to endogenous pigment, therefore we recommend you substitute an AEC or a Fast Red substrate protocol.
|IHC-Fr||1/25 - 1/50. ABC method.|
RelevanceMalignant melanoma is a malignant neoplasm of melanocytes, arising de novo or from a pre existing benign nevus, which occurs most often in the skin but also may involve other sites.
Cellular localizationMembrane; Single-pass membrane protein
- Antigen LB39 AA antibody
- Antigen SK29 AA antibody
- MART1 antibody
IHC image of ab733 staining in human melanoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab733, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ab733 staining Melanoma by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).
Overlay histogram showing MALME 3M cells stained with ab733 (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 (ab733, 1/100 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 IgG (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.
Immunofluorescence analysis of cultured Human melanoma cells, staining Melanoma with ab733.
Cells were fixed with formaldehyde and blocked with blocking reagent for 30 minutes at room temperature. Samples were incubated with primary antibody (1/25 in blocking solution) for 1 hour at 37°C. An AlexaFluor®594-conjugated donkey anti-mouse polyclonal IgG (1/200) was used as the secondary antibody.
ab733 has been referenced in 6 publications.
- Li Z et al. Robust Photodynamic Therapy Using 5-ALA-Incorporated Nanocomplexes Cures Metastatic Melanoma through Priming of CD4+CD8+ Double Positive T Cells. Adv Sci (Weinh) 6:1802057 (2019). PubMed: 30886812
- Lin H et al. Host expression of PD-L1 determines efficacy of PD-L1 pathway blockade-mediated tumor regression. J Clin Invest 128:805-815 (2018). PubMed: 29337305
- Ness C et al. Multicellular tumor spheroids of human uveal melanoma induce genes associated with anoikis resistance, lipogenesis, and SSXs. Mol Vis 23:680-694 (2017). PubMed: 29033534
- Kundu S et al. Heparanase Promotes Glioma Progression and Is Inversely Correlated with Patient Survival. Mol Cancer Res 14:1243-1253 (2016). PubMed: 27565180
- Savla R et al. Tumor-targeted responsive nanoparticle-based systems for magnetic resonance imaging and therapy. Pharm Res 31:3487-502 (2014). PubMed: 24919932
- Giordano G et al. The role of cervical smear in the diagnosis and management of extrauterine malignancies metastatic to the cervix: three case reports. Diagn Cytopathol 38:41-6 (2010). IHC-P ; Human . PubMed: 19626626