製品の概要

  • 製品名
    Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3]
    Melanoma 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [HMB45 + M2-7C10 + M2-9E3] to Melanoma
  • 由来種
    Mouse
  • アプリケーション
    適用あり: Flow Cyt, IHC-P, IHC-Frmore details
  • 種交差性
    交差種: Mouse, Human
    交差が予測される動物種: Rat
  • 免疫原

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

  • エピトープ
    HMB45 and MART-1
  • ポジティブ・コントロール
    • IHC-P: 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. This cocktail is HMB45 + two different clones of MART-1[M2-7C10] + [M2-9E3]. HMB34 isotype: IgG1/kappa and both clones of MART-1: IgG2b + IgG2b

    HMB-45 and MART-1 are coexpressed in the majority of melanomas, as well as uniquely expressed in certain cases. Thus, the HMB-45 and MART-1 cocktail is potentially more sensitive than HMB-45 or MART-1 alone. MART-1 is a cocktail of clones M2-7C10 and M2-9E3. The combination of HMB-45 and the MART-1 cocktail make this triple antibody cocktail a first-order pan melanoma screener.

    This product was changed from ascites to tissue culture supernatant on [14/07/17]. The following lots are from ascites and are still in stock as of [14/07/17] – [GR313023, GR3175007]. Lot numbers higher than [GR313023] will be from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.

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: 0.099% Sodium azide
    Constituents: 0.9% Proprietary component, 99% Water
  • 精製度
    Tissue culture supernatant
  • 一次抗体 備考
    HMB-45 and MART-1 are coexpressed in the majority of melanomas, as well as uniquely expressed in certain cases. Thus, the HMB-45 and MART-1 cocktail is potentially more sensitive than HMB-45 or MART-1 alone. MART-1 is a cocktail of clones DT101 and BC199. The combination of HMB-45 and the MART-1 cocktail make this triple antibody cocktail a first-order pan melanoma screener.
  • ポリ/モノ
    モノクローナル
  • クローン名
    HMB45 + M2-7C10 + M2-9E3
  • ミエローマ
    unknown
  • アイソタイプ
    IgG2b
  • 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 ab732 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 Use at an assay dependent concentration. PubMed: 22147606

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

 

IHC-P 1/100 - 1/200.

The staining intensity of this melanoma cocktail is enhanced by performing heat mediated antigen retrieval using a high pH buffer. 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.

IHC-Fr Use at an assay dependent concentration.

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

  • ab732 staining Melanoma in Human lymph node tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

  • ab732 staining murine melanoma metastasis tissue sections by IHC-P.  Tissue sections were paraformaldehyde fixed and blocked with 0.5% Perkin-Elmer TNB Blocking Buffer for 30 minutes at 25°C.  The primary antibody was diluted 1/50 and incubated for 18 hours at 4°C.  An Alexa Fluor 488® conjugated goat anti-mouse was used as the secondary

    Confocal image shows melanoma in magenta, vessels in red (Collagen Type IV, ab19808), and melanin in cyan. Tissue is perfusion-fixed 15µm cryostat sections mounted on slides (i.e., lightly fixed, but not paraffin embedded).

    See Abreview

参考文献

This product has been referenced in:
See all 17 Publications for this product

レビューと Q&A

1-10 of 10 Abreviews or Q&A

Answer

Thank you for contacting Abcam.
I have attached the Abcam, mouse on mouse staining protocol to see if that can give you any suggestions. If you can send me a copy of the protocol you are using, then I would be happy to see if there are any suggestions I can make.
I look forward to your reply.

Read More
Application
Immunocytochemistry
Sample
Human Cultured Cells (Human Melanoma cell line M238)
Permeabilization
No
Specification
Human Melanoma cell line M238
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative
Methanol

Rong Huang

Verified customer

投稿 Apr 16 2018

Answer

I am sorry to confirm we do not currently have fluorochrome conjugated Anti-Melanoma antibody [HMB45 + DT101 + BC199] (ab732) on our catalog.


However, you may like to consider either using a secondary antibody (https://www.abcam.com/index.html?pageconfig=productmap&cl=918) or conjugating the antibody yourself using our EasyLink antibody coniugation kits suitable for fluorochrome detection methods.

EasyLink antibody conjugation kits provide a simple and quick linking process of your antibody with the advantage of one-step antibody labeling method with no separation steps. You can find more information about our EasyLink and procedures by clicking the following link:
https://www.abcam.com/index.html?pageconfig=resource&rid=13148#1

Although we do not have EasyLink antibody conjugation kit specifically for Alexa 647, we do have EasyLink kit for dyes with similar emission and excitation wavelengths as Alexa647: APC-Cy5.5, Cy5.5 and APC-Cy7. You can find them on the table at the bottom page of the link above, where you can also find the references of the different kits we dispose (you just have to click on them to have more information). The differences between those kits is the amount of reaction allowed.

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Question
Answer

Our Melanoma Cocktail is HMB45[HMB45] + two different clones of MART-1[M2-7C10] + [M2-9E3].
HMB34 isotype: IgG1/kappa and both clones of MART-1: IgG2b + IgG2b
Epitope/antigen: HMB45 and MART-1

Read More

Answer

Thank you very much for contacting us with your enquiry.
I am sorry to hear that there has been some variability with ab732. We don't have any other recent reports of the antibody not working, and there haven't been any changes to the antibody, so I'm not sure if this is a general problem or if it a faulty vial. The new vial that you have ordered is from a different production batch than the problematic one, so the results may improve using this vial.
Regarding the problematic vial, we do fully guarantee this antibody to work in the tested species and applications listed on the datasheet. Could you send me some details about the sample and protocol used with this antibody? Were the same samples and protocol used with the old and new vials? Do you have your order number for the vial that didn't work? I can arrange a credit or refund if you would like.
I look forward to hearing from you. Please let me know if you have further questions or if there is anything else that we can do for you.

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Answer

Thank you for your enquiry. I am pleased to provide the following suggested positive controls for these antibodies. I have mainly chosen tissue that has been tested succesfully and shown in an image on the datasheets: 1.Anti-ErbB 2 antibody (ab2428) Human breast cancer 2.Anti-Progesterone Receptor antibody [SP2] (ab16661) Human breast carcinoma 3.Anti-Estrogen Receptor antibody [SP1] (ab16660) Human breast carcinoma 4.Anti-Myeloperoxidase antibody (ab45977) Human spleen tissue 5.Anti-TdT antibody (ab85148) Human testis 6.Anti-TMEM16A antibody (ab53212) Human liver 7.Anti-NSE antibody [EPR3377] (ab79757) Human brain tissue 8.Anti-CD23 antibody [EPR3617(IHC)] (ab92495) Human tonsil 9.Anti-CD10 antibody [56C6] (ab951) Lymph node tissue or human breast microepithelium 10.Anti-Melanoma antibody [HMB45 + DT101 + BC199] (ab732) Murine melanoma metastasis tissue 11.Anti-CD79a antibody [HM47/A9] (ab3121) Human Lymphoid tissue, tonsil 12.Anti-CD45RO antibody [UCHL-1] (ab86080) Human Tonsil, Thymus 13.Anti-CD45 antibody (ab10559) Human Tonsil tissue 14.Anti-CD30 antibody [CON6D/ C2] - BSA and Azide free (ab76518) Human Hodgekins lymphoma tissue 15.Anti-CD15 antibody [(Workshop IV) (BRA-4F1) our designation 238)] (ab115993) Human Hodgekins lymphoma tissue Human spleen I am providing a list ofslides that we sell which I hope will be helpful to you. I can recommend to review the online datasheet for more details. Human brain: ab4305Brain (human): hippocampus normal tissue slides https://www.abcam.com/index.html?datasheet=4305 (or use the following: https://www.abcam.com/index.html?datasheet=4305). Human lymph node: ab4350Lymph node (human) normal tissue slides IHC-P, ISH https://www.abcam.com/index.html?datasheet=4350 (or use the following: https://www.abcam.com/index.html?datasheet=4350). Human testis: ab4373Testis (human) normal tissue slides IHC-P, ISH https://www.abcam.com/index.html?datasheet=4373 (or use the following: https://www.abcam.com/index.html?datasheet=4373). Human liver: ab4348Liver (human) normal tissue slides IHC-P, ISH https://www.abcam.com/index.html?datasheet=4348 (or use the following: https://www.abcam.com/index.html?datasheet=4348). Human spleen: ab4372Spleen (human) normal tissue slides IHC-P, ISH https://www.abcam.com/index.html?datasheet=4372 (or use the following: https://www.abcam.com/index.html?datasheet=4372). I am sorry we do not have slides forthe following: Human tonsil Hodgekins lymphoma tissue Mouse melenoma tissue I hope this will be helpful to you. If you have any further questions,please do not hesitate to contact us.

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Answer

Thank you for your patience while I was looking into this. I have heard back from the lab and they have told me that ab732 was tested using the DIVA decloaking agent (http://biocare.net/wp-content/uploads/DV2004X.pdf) which is at pH.6.2. Also for the refrence of the antibody being tested in IHC paraffin embedded tissue, the only ones I can find of the antibody beind used on mouse tissue and/or on paraffin embedded sections are; Azarova AM et al. Roles of DNA topoisomerase II isozymes in chemotherapy and secondary malignancies. Proc Natl Acad Sci U S A 104:11014-9 (2007). Jungbluth AA et al. A103: An anti-melan-a monoclonal antibody for the detection of malignant melanoma in paraffin-embedded tissues. Am J Surg Pathol 22:595-602 (1998). If there is anything else I can help you with, please let me know.  

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Question
Answer

Thank you for contacting Abcam. The 3 clones in the ab732 cocktail do recognize mouse tissue. The IHCP protocol that was used to test the antibody was optimized to human tissue and the following protocol was used: Pretreatment Protocol Heat Retrieval Method: Retrieve sections under pressure using BIOCARE's Decloaking Chamber, followed by a wash in distilled water. Alternatively, steam tissue sections for 45-60 minutes. Allow solution to cool for 20 minutes then wash in distilled water. Protein Block: Incubate for 10-15 minutes at RT with BIOCARE's Background Sniper. Primary Antibody: Incubate for 30 minutes at RT. Probe: Incubate for 10 minutes at RT with a Probe. Chromogen: Incubate for 5 minutes at RT when using BIOCARE's DA Polymer : Incubate for 10 minutes at RT with a Polymer. If there is anything else I can help with, please let me know.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (murine melanoma metastasis)
Specification
murine melanoma metastasis
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No
Blocking step
(agent) for 30 minute(s) · Concentration: 0.5% · Temperature: 25°C

Abcam user community

Verified customer

投稿 Jan 07 2008

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

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