RecombinantRabMAb

Recombinant Anti-Melanoma gp100 antibody [EPR4864] (ab137062)

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Western blot - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
  • Western blot - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
  • Anti-Melanoma gp100 antibody [EPR4864] (ab137062)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4864] to Melanoma gp100
  • Suitable for: WB, IHC-P
  • Reacts with: Human

Overview

  • Product name

    Anti-Melanoma gp100 antibody [EPR4864]
    See all Melanoma gp100 primary antibodies

  • Description

    Rabbit monoclonal [EPR4864] to Melanoma gp100

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: ICC or IP

  • Species reactivity

    Reacts with: Human

  • Immunogen

    within Human Melanoma gp100 aa 600 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P40967

  • Positive control

    • Human melanoma tissue lysate; Human melanoma tissue ;Placenta tissue; Normal skin tissue;Normal kidney tissue.

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab137062 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
WB 1/1000 - 1/10000. Detects a band of approximately 100, 26 kDa (predicted molecular weight: 70 kDa).

For unpurified, use 1/1000 - 1/2000.
IHC-P 1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

For unpurified, use 1/50 - 1/100.

See IHC antigen retrieval protocols.
  • Application notes
    Is unsuitable for ICC or IP.

    • 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

      • Western blot - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
        Western blot - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
        Anti-Melanoma gp100 antibody [EPR4864] (ab137062) at 1/10000 dilution (purified) + Human melanoma tissue lysate at 10 µg

        Secondary
        HRP goat anti-rabbit (H+L) at 1/1000 dilution

        Predicted band size: 70 kDa
        Observed band size: 100 kDa
        why is the actual band size different from the predicted?
        Additional bands at: 26 kDa. We are unsure as to the identity of these extra bands.



        Blocking buffer: 5% NFDM/TBST

        Dilution buffer: 5% NFDM/TBST

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)

        Immunohistochemical staining of paraffin embedded human melanoma with purified ab137062 at a working dilution of 1 in 400. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

      • Western blot - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
        Western blot - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
        Anti-Melanoma gp100 antibody [EPR4864] (ab137062) at 1/1000 dilution (unpurified) + Human melanoma tissue lysate at 10 µg

        Secondary
        HRP labelled Goat anti-Rabbit IgG at 1/2000 dilution

        Predicted band size: 70 kDa

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)

        Immunohistochemical staining of paraffin embedded human melanoma with unpurified ab137062 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)

        Immunohistochemical analysis using unpurified ab137062 showing negative staining in placenta tissue.

        Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)

        Immunohistochemical analysis using unpurifed ab137062 showing negative staining in normal skin tissue.

        Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)

        Immunohistochemical analysis using unpurified ab137062 showing negative staining in normal kidney tissue.

        Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EPR4864] (ab137062)

        Immunohistochemical analysis using unpurified ab137062 showing negative staining in normal stomach tissue.

        Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

      • Anti-Melanoma gp100 antibody [EPR4864] (ab137062)
        Anti-Melanoma gp100 antibody [EPR4864] (ab137062)

      References (6)

      Publishing research using ab137062? Please let us know so that we can cite the reference in this datasheet.

      ab137062 has been referenced in 6 publications.

      • Graham M  et al. Repeat domain-associated O-glycans govern PMEL fibrillar sheet architecture. Sci Rep 9:6101 (2019). PubMed: 30988362
      • Yi WJ  et al. Degraded melanocores are incompetent to protect epidermal keratinocytes against UV damage. Cell Cycle 17:844-857 (2018). PubMed: 29623762
      • McGill TJ  et al. Allogeneic iPSC-Derived RPE Cell Graft Failure Following Transplantation Into the Subretinal Space in Nonhuman Primates. Invest Ophthalmol Vis Sci 59:1374-1383 (2018). Macaque monkey . PubMed: 29625461
      • Regazzetti C  et al. Melanocytes Sense Blue Light and Regulate Pigmentation through Opsin-3. J Invest Dermatol 138:171-178 (2018). PubMed: 28842328
      • Zhou D  et al. A novel P53/POMC/Gas/SASH1 autoregulatory feedback loop activates mutated SASH1 to cause pathologic hyperpigmentation. J Cell Mol Med 21:802-815 (2017). WB, IHC-P ; Human . PubMed: 27885802
      • McGill TJ  et al. Long-Term Efficacy of GMP Grade Xeno-Free hESC-Derived RPE Cells Following Transplantation. Transl Vis Sci Technol 6:17 (2017). IHC-P ; Rat . PubMed: 28626601

      Customer reviews and Q&As

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      Immunohistochemistry (Frozen sections) abreview for Anti-Melanoma gp100 antibody [EPR4864]

       Excellent
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Frozen sections)
      Blocking step
      Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 22°C
      Sample
      Human Tissue sections (Melanoma)
      Specification
      Melanoma
      Permeabilization
      Yes - 0.5%
      Fixative
      Paraformaldehyde
      Read More

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      Verified customer

      Submitted May 28 2014

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