Anti-Melanoma gp100 antibody [HMB45] (ab787)

Reviews (2)Q&A (3)References (24)

Overview

  • Product name

    Anti-Melanoma gp100 antibody [HMB45]
    See all Melanoma gp100 primary antibodies

  • Description

    Mouse monoclonal [HMB45] to Melanoma gp100

  • Host species

    Mouse

  • Tested applications

    Suitable for: Flow Cyt, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Human
    Does not react with: Rat, Dog

  • Immunogen

    Tissue, cells or virus corresponding to Human Melanoma. BALB/C mice were injected with extract of pigmented malanoma metastases from lymph nodes.
    Database link: P40967

  • Positive control

    • IHC-P: Human melanoma and testis tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab787 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 1/10.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P Use a concentration of 0.5 - 1 µg/ml.

Incubate for 30 minutes at room temperature. Staining of formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0, for 10-20 min followed by cooling at RT for 20 minutes
ICC/IF Use a concentration of 0.5 - 1 µg/ml. PubMed: 19841138

Target

  • Function

    Plays a central role in the biogenesis of melanosomes. Involved in the maturation of melanosomes from stage I to II. The transition from stage I melanosomes to stage II melanosomes involves an elongation of the vesicle, and the appearance within of distinct fibrillar structures. Release of the soluble form, ME20-S, could protect tumor cells from antibody mediated immunity.

  • Tissue specificity

    Preferentially expressed in melanomas. Some expression was found in dysplastic nevi. Not found in normal tissues nor in carcinomas. Normally expressed at low levels in quiescent adult melanocytes but overexpressed by proliferating neonatal melanocytes and during tumor growth.

  • Sequence similarities

    Belongs to the PMEL/NMB family.
    Contains 1 PKD domain.

  • Domain

    The RPT domain is essential for the generation of the fibrillar matrix of melanosomes.
    The lumenal domain is necessary for correct processing and trafficking to melanosomes.

  • Post-translational
    modifications

    A small amount of P1/P100 (major form) undergoes glycosylation to yield P2/P120 (minor form). P2 is cleaved by a furin-like proprotein convertase (PC) in a pH-dependent manner in a post-Golgi, prelysosomal compartment into two disulfide-linked subunits: a large lumenal subunit, M-alpha/ME20-S, and an integral membrane subunit, M-beta. Despite cleavage, only a small fraction of M-alpha is secreted, whereas most M-alpha and M-beta remain associated with each other intracellularly. M-alpha is further processed to M-alpha N and M-alpha C. M-alpha C further undergoes processing to yield M-alpha C1 and M-alpha C3 (M-alpha C2 in the case of PMEL17-is or PMEL17-ls). Formation of intralumenal fibrils in the melanosomes requires the formation of M-alpha that becomes incorporated into the fibrils. Stage II melanosomes harbor only Golgi-modified Pmel17 fragments that are derived from M-alpha and that bear sialylated O-linked oligosaccharides.
    N-glycosylated. O-glycosylated; contains sialic acid.

  • Cellular localization

    Secreted and Endoplasmic reticulum membrane. Golgi apparatus. Melanosome. Endosome > multivesicular body. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. Localizes predominantly to intralumenal vesicles (ILVs) within multivesicular bodies. Associates with ILVs found within the lumen of premelanosomes and melanosomes and particularly in compartments that serve as precursors to the striated stage II premelanosomes.
  • Information by UniProt

  • Database links

    • Alternative names

      • 95 kDa melanocyte specific secreted glycoprotein antibody
      • 95 kDa melanocyte-specific secreted glycoprotein antibody
      • D12S53E antibody
      • gp100 antibody
      • M-beta antibody
      • ME20 antibody
      • ME20 M/ME20 S antibody
      • ME20-M antibody
      • ME20-S antibody
      • ME20M antibody
      • ME20M/ME20S antibody
      • ME20S antibody
      • Melanocyte lineage specific antigen GP100 antibody
      • Melanocyte protein mel 17 antibody
      • Melanocyte protein Pmel 17 antibody
      • Melanocyte protein Pmel 17 precursor antibody
      • Melanocytes lineage-specific antigen GP100 antibody
      • Melanoma associated ME20 antigen antibody
      • Melanoma gp100 antibody
      • Melanoma-associated ME20 antigen antibody
      • Melanosomal matrix protein 17 antibody
      • Melanosomal matrix protein17 antibody
      • P1 antibody
      • p100 antibody
      • p26 antibody
      • PMEL 17 antibody
      • PMEL antibody
      • PMEL_HUMAN antibody
      • PMEL17 antibody
      • Premelanosome protein antibody
      • Secreted melanoma-associated ME20 antigen antibody
      • SI antibody
      • SIL antibody
      • SILV antibody
      • Silver (mouse homolog) like antibody
      • Silver homolog antibody
      • Silver locus protein homolog antibody
      • Silver, mouse, homolog of antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)

      Formalin-fixed, paraffin-embedded human melanoma tissue stained with ab787 at 1 μg/ml in immunohistochemical analysis. 

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)

      ab787 - immunohistochemistry

      Formalin fixed paraffin embedded human melanoma stained with ab787, using ABC and AEC chromogen.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)

      Formalin-fixed, paraffin-embedded human testis tissue stained with ab787 at 1 μg/ml in immunohistochemical analysis. 

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [HMB45] (ab787)This image is courtesy of an Abreview submitted by Dr. Hongwei Shao
      ab787 staining Melanoma in the human Melanoma cell line WM3248 xenopraph by IHC-P (formaldehyde-fixed paraffin-embedded sections). Tissue samples were fixed with formaldehyde; permeabilized with 0.1% Triton X-100 and blocked with 100% Dakocytomation X0909 for 1 hour at room temperature; antigen retrival was by heat mediation in Citric buffer (pH6). The sample was incubated with primary antibody (1/25) at 4°C for 18 hours. An HRP-conjugated Goat polyclonal to mouse IgG (1/100) was used as secondary antibody.

      See Abreview

    • Flow Cytometry - Anti-Melanoma gp100 antibody [HMB45] (ab787)
      Flow Cytometry - Anti-Melanoma gp100 antibody [HMB45] (ab787)
      Overlay histogram showing Malme-3 cells stained with ab787 (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. The cells were then incubated with the antibody (ab787, 1/10 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Malme-3 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

    References

    This product has been referenced in:

    • Zhou X  et al. Melanocyte Chitosan/Gelatin Composite Fabrication with Human Outer Root Sheath-Derived Cells to Produce Pigment. Sci Rep 9:5198 (2019). Read more (PubMed: 30914712) »
    • Fang Y  et al. Exosomal miRNA-106b from cancer-associated fibroblast promotes gemcitabine resistance in pancreatic cancer. Exp Cell Res 383:111543 (2019). Read more (PubMed: 31374207) »
    See all 24 Publications for this product

    Publishing research using ab787? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-5 of 5 Abreviews or Q&A

    Question

    I was wondering how long the product is stable for 4 degrees? This is not stated on the certificate of compliance.

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    Answer



    We don't have strict guidelines for length of storage for antibodies. In general, for antibody products stored at 4 degrees C, we would expect them to be stable for at least one year from date of purchase.

    Read More

    Question

    related to CCE 4314538.

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    Answer

    Merci de nous avoir contactés; ma collègue m'a fait part de votre appel.

    Je suis heureux de savoir que l'anticorps a finalement fonctionné en cytométrie en flux. J'ai compris que vous deviez utiliser beaucoup d'anticorps pour obtenir des résultats satisfaisants sur 300.000 cellules. J'ai donc mis en place l'envoi d'une unité gratuite d'ab787 pour que vous puissiez terminer vos travaux avec cet anticorps.

    Le numéro de la commande est ****.

    N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

    Read More

    Question

    Product code: 787
    Inquiry: Hi, I'm looking at this product: Anti-Melanoma antibody [HMB45] (ab787) Do you have a protein/peptide that this antibody recognizes? I need to use the protein/peptide in my experiment as a positive control. Otherwise, are you able to recommend an antibody/antigen pair that can be used in this way? It doesn't have to be HMB45 strictly any human melanoma marker will do. I've been looking also at S100 in your catalog but I don't know which antibodies/proteins are compatible. Thanks.

    Read More
    Answer

    Thank you for contacting us.

    Unfortunately, the blocking peptide for ab787 antibody is not available for purchase. We have ˜25 anti melanoma antibodies available but we do not have antigen available for any one of these however if you could select few as per your requirement then I will check with our collaborator laboratory if they can supply the peptide as special request.

    We do have antibody / antigen pair for S100 protein. Please check the catalogue number ab14688 and ab30380.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

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

     Average
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Melanoma cell line WM3248 xenopraph)
    Specification
    Melanoma cell line WM3248 xenopraph
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric buffer pH6.0
    Permeabilization
    Yes - 0.1% triton x-100
    Blocking step
    Dakocytomation x0909 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: RT°C
    Read More

    Hongwei Shao

    Verified customer

    Submitted Feb 25 2010

    Western blot abreview for Anti-Melanoma antibody [HMB45]

     Below Average
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - other (transfected HeLa)
    Loading amount
    20 µg
    Specification
    transfected HeLa
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Dr. Guillaume Van Niel

    Verified customer

    Submitted Jul 18 2007

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