Anti-Melanoma antibody [HMB45 + DT101 + BC199 + T311] (ab733)

Overview

  • Product nameAnti-Melanoma antibody [HMB45 + DT101 + BC199 + T311]
    See all Melanoma primary antibodies
  • Description
    Mouse monoclonal [HMB45 + DT101 + BC199 + 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 reactivity
    Reacts with: Human
  • Immunogen

    HMB45 - Pigmented melanoma metastases from LN MART-1 - Recombinant human MART-1 protein Tyrosinase - Recombinant tyrosinase protein

  • Positive control
    • Metastatic melanoma in lymph node.
  • General notes


    The 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.).

Properties

  • FormLiquid
  • 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 bufferPreservative: Proprietary, not sodium azide or thimerosal.
    Constituents: PBS, Tissue Culture Supernatant. pH 7.3; 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.).
  • ClonalityMonoclonal
  • Clone numberHMB45 + DT101 + BC199 + T311
  • Myelomaunknown
  • IsotypeIgG
  • Light chain typekappa
  • Research areas

Associated products

Applications

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.

Application Abreviews Notes
Flow Cyt 1/100.
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. ABC method The HMB-45 portion of this antibody will stain with citrate buffer, pH 6.0; however, data shows the MART-1 portion of this cocktail may be negative unless a high pH buffer is used. We therefore, strongly recommend that you do not use citrate buffer for this antibody cocktail. 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.

Target

  • 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
  • Database links
  • Alternative names
    • Antigen LB39 AA antibody
    • Antigen SK29 AA antibody
    • MART1 antibody
    • melan A antibody
    • Melan A protein antibody
    • Melanoma antigen recognized by T-cells 1 antibody
    • Monophenol monooxygenase antibody
    • Tumor rejection antigen AB antibody
    • tyrosinase antibody
    see all

Anti-Melanoma antibody [HMB45 + DT101 + BC199 + T311] images

  • 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.
  • 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.

  • 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.

    See Abreview

References for Anti-Melanoma antibody [HMB45 + DT101 + BC199 + T311] (ab733)

This product has been referenced in:
  • 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 . Read more (PubMed: 19626626) »

See 1 Publication for this product

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cultured Cells (melanoma cell line)
Specification melanoma cell line
Fixative Formaldehyde
Permeabilization No
Blocking step Dakocytomation # X0909 as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: RT°C
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Hongwei Shao

Verified customer

Submitted Jan 18 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"