RabMAb

HRP Anti-Melanoma gp100 antibody [EP4863(2)] (ab205469)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-Melanoma gp100 antibody [EP4863(2)] (ab205469)
  • Western blot - HRP Anti-Melanoma gp100 antibody [EP4863(2)] (ab205469)

Key features and details

  • HRP Rabbit monoclonal [EP4863(2)] to Melanoma gp100
  • Suitable for: IHC-P, WB
  • Reacts with: Mouse, Human
  • Conjugation: HRP
  • Isotype: IgG

Overview

  • Product name

    HRP Anti-Melanoma gp100 antibody [EP4863(2)]
    See all Melanoma gp100 primary antibodies

  • Description

    HRP Rabbit monoclonal [EP4863(2)] to Melanoma gp100

  • Host species

    Rabbit

  • Conjugation

    HRP

  • Tested applications

    Suitable for: IHC-P, WBmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Synthetic peptide within Human Melanoma gp100 aa 1-100. The exact sequence is proprietary.
    Database link: P40967

  • Positive control

    • WB: B16-F0 cell lysate, human melanoma tissue lysate. IHC-P: FFPE Human skin melanoma.

  • General notes

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

    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 ab205469 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 1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 70 kDa).

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) - HRP Anti-Melanoma gp100 antibody [EP4863(2)] (ab205469)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-Melanoma gp100 antibody [EP4863(2)] (ab205469)

      IHC image of gp100 staining in a section of formalin-fixed paraffin-embedded Human Skin Melanoma*, performed on a Leica BONDTM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab205469, 1/50 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • Western blot - HRP Anti-Melanoma gp100 antibody [EP4863(2)] (ab205469)
      Western blot - HRP Anti-Melanoma gp100 antibody [EP4863(2)] (ab205469)
      All lanes : HRP Anti-Melanoma gp100 antibody [EP4863(2)] (ab205469) at 1/1000 dilution

      Lane 1 : B16-F0 (Mouse) Whole Cell Lysate
      Lane 2 : Melanoma (Human) Whole Cell Lysate - tumor tissue

      Lysates/proteins at 10 µg per lane.

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 70 kDa
      Observed band size: 80 kDa
      why is the actual band size different from the predicted?


      Exposure time: 8 minutes


      This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab205469 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

    References (0)

    Publishing research using ab205469? Please let us know so that we can cite the reference in this datasheet.

    ab205469 has not yet been referenced specifically in any publications.

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