Anti-Melanoma 抗体 [HMB45 + M2-7C10 + M2-9E3 + T311] (ab733)

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製品の概要

  • 製品名
    Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3 + T311]
    Melanoma 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [HMB45 + M2-7C10 + M2-9E3 + T311] to Melanoma
  • 由来種
    Mouse
  • 特異性
    The 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.
  • アプリケーション
    適用あり: Flow Cyt, ICC/IF, IHC-P, IHC-Frmore details
  • 種交差性
    交差種: Human
  • 免疫原

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

  • ポジティブ･コントロール
    • Metastatic melanoma in lymph node.
  • 特記事項

    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) 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.).

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファー
    Preservative: Proprietary, not sodium azide or thimerosal.
    Constituents: PBS, Tissue Culture Supernatant. pH 7.3; Protein carrier.
  • Concentration information loading...
  • 一次抗体 備考
    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.).
  • ポリ/モノ
    モノクローナル
  • クローン名
    HMB45 + M2-7C10 + M2-9E3 + T311
  • ミエローマ
    unknown
  • アイソタイプ
    IgG
  • 軽鎖の種類
    kappa
  • 研究分野

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

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.

ターゲット情報

  • 関連性
    Malignant 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.
  • 細胞内局在
    Membrane; Single-pass membrane protein
  • 参照データベース
    • 別名
      • 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

    画像

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3 + T311] (ab733)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3 + T311] (ab733)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3 + T311] (ab733)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3 + T311] (ab733)

      ab733 staining Melanoma by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

    • Flow Cytometry - Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3 + T311] (ab733)
      Flow Cytometry - Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3 + T311] (ab733)
      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.
    • Immunocytochemistry/ Immunofluorescence - Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3 + T311] (ab733)
      Immunocytochemistry/ Immunofluorescence - Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3 + T311] (ab733)This image is courtesy of an Abreview submitted by Hongwei Shao
      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

    参考文献

    This product has been referenced in:
    • 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). Read more (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). Read more (PubMed: 29033534) »
    See all 5 Publications for this product

    ab733 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    レビューと Q&A

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    1-2 of 2 Abreviews or Q&A

    Question

    Phone call requesting concentration of antibody.

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    Answer

    Thank you for contacting us and your interest in our products.

    I am sorry for the delay in getting back to you. I can now confirm that the anti-Melanoma antibody [HMB45 + DT101 + BC199 + T311] (ab733) is composed of a combination of HMB45, MART-1 (DT101 + BC199) and Tyrosinase (T311) make this a quadruple antibody cocktail, a first-order pan melanoma screener. The concentration of each of the antibodies in the lot which you have received is as follows:

    HMB45 is 8.9µg/mL;
    MART-1 [m2-E93] is 22.2µg/mL;
    Melan A [M2-7C10] is 20µg/mL;
    Tyrosinase is 27µg/mL;

    I hope this has been helpful. If I can be of any further assistance, please do not hesitate to let me know.

    Read More

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Melanoma antibody [HMB45 + DT101 + BC199 + T311]

     Excellent
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    abreview image
    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
    Read More

    Hongwei Shao

    Verified customer

    投稿 Jan 18 2013

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

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