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Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3] (ab732)

  • Datasheet
  • SDS
Reviews (2)Q&A (8)References (21)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3] (ab732)

    Key features and details

    • Mouse monoclonal [HMB45 + M2-7C10 + M2-9E3] to Melanoma
    • Suitable for: IHC-P
    • Reacts with: Mouse, Human
    • Isotype: IgG2b

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    Overview

    • Product name

      Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3]
      See all Melanoma primary antibodies
    • Description

      Mouse monoclonal [HMB45 + M2-7C10 + M2-9E3] to Melanoma
    • Host species

      Mouse
    • Tested applications

      Suitable for: IHC-Pmore details
    • Species reactivity

      Reacts with: Mouse, Human
      Predicted to work with: Rat
    • Immunogen

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

    • Epitope

      HMB45 and MART-1
    • Positive control

      • IHC-P: Metastatic melanoma in lymph node.
    • General notes

      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.

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

      One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

      Learn more here.

    Properties

    • Form

      Liquid
    • Storage instructions

      Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
    • Storage buffer

      Preservative: 0.099% Sodium azide
      Constituents: 0.9% Proprietary component, 99% Water
    • Concentration information loading...
    • Purity

      Tissue culture supernatant
    • Primary antibody notes

      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.
    • Clonality

      Monoclonal
    • Clone number

      HMB45 + M2-7C10 + M2-9E3
    • Myeloma

      unknown
    • Isotype

      IgG2b
    • Light chain type

      kappa
    • Research areas

      • Tags & Cell Markers
      • Cell Type Markers
      • Tumor Associated
      • Cancer
      • Tumor biomarkers
      • Other

    Associated products

    • Compatible Secondaries

      • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
      • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Isotype control

      • Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)

    Applications

    The Abpromise guarantee

    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.

    Application Abreviews Notes
    IHC-P (1)
    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.

    Notes
    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.

    Target

    • Relevance

      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.
    • Cellular localization

      Membrane; Single-pass membrane protein
    • Database links

      • Entrez Gene: 77836 Mouse
      • Entrez Gene: 293890 Rat
      • Omim: 605513 Human
      • SwissProt: Q16655 Human
      • SwissProt: Q2TA50 Mouse
      • 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
        see all

      Images

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3] (ab732)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3] (ab732)

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

      Protocols

      • Immunohistochemistry protocols

      Click here to view the general protocols

      Datasheets and documents

      • SDS download

      • Datasheet download

        Download

      References (21)

      Publishing research using ab732? Please let us know so that we can cite the reference in this datasheet.

      ab732 has been referenced in 21 publications.

      • Golan T  et al. Adipocytes sensitize melanoma cells to environmental TGF-ß cues by repressing the expression of miR-211. Sci Signal 12:N/A (2019). PubMed: 31337739
      • Li H  et al. YAP/TAZ Activation Drives Uveal Melanoma Initiation and Progression. Cell Rep 29:3200-3211.e4 (2019). PubMed: 31801083
      • Han N  et al. Enhanced IL-34 expression in Nivolumab-resistant metastatic melanoma. Inflamm Regen 38:3 (2018). PubMed: 29515691
      • Malcov-Brog H  et al. UV-Protection Timer Controls Linkage between Stress and Pigmentation Skin Protection Systems. Mol Cell 72:444-456.e7 (2018). PubMed: 30401431
      • Yanaki M  et al. In Vivo Antitumor Effects of MK615 Led by PD-L1 Downregulation. Integr Cancer Ther 17:646-653 (2018). PubMed: 29665734
      View all Publications for this product

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      1-10 of 10 Abreviews or Q&A

      Question


      My lab ordered an antibody(Cat No: ab732) from Abcam. It is mouse anti-mouse Melanoma antibody [HMB45 + DT101 + BC199] (ab732). My sample is from mouse. When I did the Immunohistochical staining, the background is very strong. I used goat anti-mouse Peroxidase affinipure Fab goat anti mouse IgG (H+L) as secondary antibody from Jackson immunoresearch and used 3,3-diaminobenzidine substrate system. Before added the primary antibody, I have block with 5%BSA solution. Do you have any idea to block this mouse ant-mouse antibody's background? Thanks

      Read More

      Abcam community

      Verified customer

      Asked on Mar 23 2012

      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

      Abcam Scientific Support

      Answered on Mar 23 2012

      Immunocytochemistry abreview for Anti-Melanoma antibody [HMB45 + M2-7C10 + M2-9E3]

      Inconclusive
      Abreviews
      Abreviews
      abreview image
      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
      Read More

      Rong Huang

      Verified customer

      Submitted Apr 16 2018

      Question



      Inquiry: Hi, Is it possible to get 732 Anti-Melanoma antibody [HMB45 + DT101 + BC199] stained with a fluorochrome, eg Alexa 647?

      Read More

      Abcam community

      Verified customer

      Asked on Sep 17 2013

      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.

      Read More

      Abcam Scientific Support

      Answered on Sep 17 2013

      Question

      ab732: What is the clonality of each antibody in the mixture? Are they all IgG2b.

      Read More

      Abcam community

      Verified customer

      Asked on Feb 26 2013

      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

      Abcam Scientific Support

      Answered on Feb 26 2013

      Question

      Inquiry: We just placed an order for ab732 (HMB45 mouse monoclonal antibody). The order reference number is #***. The reason we are ordering it was our previous batch was not giving us any signal, though the batch before that was very effective. Have there been any issues with this particular antibody recently or any changes in protocol? Thank you for your time

      Read More

      Abcam community

      Verified customer

      Asked on Dec 05 2012

      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.

      Read More

      Abcam Scientific Support

      Answered on Dec 05 2012

      Question

      Dear Sir/Madam Please kindly recommend the abcam slide control for IHC-P application for this antibodies. 1. Anti-ErbB 2 antibody (ab2428) 2. Anti-Progesterone Receptor antibody [SP2] (ab16661) 3. Anti-Estrogen Receptor antibody [SP1] (ab16660) 4. Anti-Myeloperoxidase antibody (ab45977) 5. Anti-TdT antibody (ab85148) 6. Anti-TMEM16A antibody (ab53212) 7. Anti-NSE antibody [EPR3377] (ab79757) 8. Anti-CD23 antibody [EPR3617(IHC)] (ab92495) 9. Anti-CD10 antibody [56C6] (ab951) 10. Anti-Melanoma antibody [HMB45 + DT101 + BC199] (ab732) 11. Anti-CD79a antibody [HM47/A9] (ab3121) 12. Anti-CD45RO antibody [UCHL-1] (ab86080) 13. Anti-CD45 antibody (ab10559) 14. Anti-CD30 antibody [CON6D/ C2] - BSA and Azide free (ab76518) 15. Anti-CD15 antibody [(Workshop IV) (BRA-4F1) our designation 238)] (ab115993) Best regards

      Read More

      Abcam community

      Verified customer

      Asked on Jan 09 2012

      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/Brain-human-hippocampus-normal-tissue-slides-ab4305.html (or use the following: https://www.abcam.com/Brain-human-hippocampus-normal-tissue-slides-ab4305.html). Human lymph node: ab4350Lymph node (human) normal tissue slides IHC-P, ISH https://www.abcam.com/Lymph-node-human-normal-tissue-slides-ab4350.html (or use the following: https://www.abcam.com/Lymph-node-human-normal-tissue-slides-ab4350.html). Human testis: ab4373Testis (human) normal tissue slides IHC-P, ISH https://www.abcam.com/Testis-human-normal-tissue-slides-ab4373.html (or use the following: https://www.abcam.com/Testis-human-normal-tissue-slides-ab4373.html). Human liver: ab4348Liver (human) normal tissue slides IHC-P, ISH https://www.abcam.com/Liver-human-normal-tissue-slides-ab4348.html (or use the following: https://www.abcam.com/Liver-human-normal-tissue-slides-ab4348.html). Human spleen: ab4372Spleen (human) normal tissue slides IHC-P, ISH https://www.abcam.com/Spleen-human-normal-tissue-slides-ab4372.html (or use the following: https://www.abcam.com/Spleen-human-normal-tissue-slides-ab4372.html). 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.

      Read More

      Abcam Scientific Support

      Answered on Jan 09 2012

      Question

      Dear John, Thank you for this information - but I'm afraid it is a bit vague.  If these Abs are known to react with murine paraffin embedded tissue sections - will you please send me the reference? Also- it is not clear what pH is used during antigen retrieval - pH10 or pH6.5 citrate?

      Read More

      Abcam community

      Verified customer

      Asked on Nov 09 2011

      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.  

      Read More

      Abcam Scientific Support

      Answered on Nov 09 2011

      Question

      Do all 3 clones recognize mouse tissue? Also what IHC-P protcol did you use?

      Read More

      Abcam community

      Verified customer

      Asked on Nov 08 2011

      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.

      Read More

      Abcam Scientific Support

      Answered on Nov 08 2011

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Melanoma antibody [HMB45 + DT101 + BC199]

      Good
      Abreviews
      Abreviews
      abreview image
      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
      Read More

      Abcam user community

      Verified customer

      Submitted Jan 07 2008

      Question

      hi, I have question about ab732,Melanoma antibody [HMB45 + DT101 + BC199] (ab732) . Data sheet says that "For formalin fixed paraffin embedded sections, 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. " What does it mean. Do i have to perform Ag retrieval or not, if yes than which buffer I have to use. Citrate ph6.0 or EDTA, pH 8.0 and what concentration of EDTA, how long I have to heat. I check protocol on your website for EDTA it did not say anything Thanks

      Read More

      Abcam community

      Verified customer

      Asked on Nov 02 2005

      Answer

      Thank you for your enquiry. You would not want to use the citrate buffer for antigen retrieval on your samples when using this antibody cocktail. You can use 1mM EDTA pH 8.0, and you should determine the optimal heating time for your sample. A good place to start would be 10-20 minutes. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

      Read More

      Abcam Scientific Support

      Answered on Nov 03 2005

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com

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