Anti-Melanoma gp100 antibody [HMB45] (ab787)

Abreviews (2)Q&A (3)Specific References (12)

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:
    • Liu W  et al. Hepatic epithelioid angiomyolipoma is a rare and potentially severe but treatable tumor: A report of three cases and review of the literature. Oncol Lett 11:3669-3675 (2016). IHC ; Human . Read more (PubMed: 27313680) »
    • Zhu L  et al. Surgical monotherapy may be a suitable therapeutic strategy for advanced collecting (Bellini) duct carcinoma: A case report and literature review. Exp Ther Med 12:1181-1184 (2016). IHC-P ; Human . Read more (PubMed: 27446340) »

    See all 12 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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    5 reviews or Q&A

    10
    Votes: Highest First

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



    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.

    related to CCE 4314538.

    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'anti...

    Read More

    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...

    Read More

    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 se...

    Read More

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

     Average
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    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
    abreview image
    Username

    Hongwei Shao

    Verified customer

    Submitted Feb 25 2010

    Western blot abreview for Anti-Melanoma antibody [HMB45]

     Below Average
    Abreviews
    Abcam has not validated the combination of species/application used in this Abreview.
    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
    Username

    Dr. Guillaume Van Niel

    Verified customer

    Submitted Jul 18 2007

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

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