RabMAb

Alexa Fluor® 647 Anti-Melanoma gp100 antibody [EP4863(2)] (ab246730)

Submit a review Submit a question
Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Melanoma gp100 antibody [EP4863(2)] (ab246730)
  • Flow Cytometry - Alexa Fluor® 647 Anti-Melanoma gp100 antibody [EP4863(2)] (ab246730)

Key features and details

  • Alexa Fluor® 647 Rabbit monoclonal [EP4863(2)] to Melanoma gp100
  • Suitable for: Flow Cyt, ICC/IF
  • Reacts with: Mouse
  • Conjugation: Alexa Fluor® 647. Ex: 652nm, Em: 668nm
  • Isotype: IgG

Overview

  • Product name

    Alexa Fluor® 647 Anti-Melanoma gp100 antibody [EP4863(2)]
    See all Melanoma gp100 primary antibodies

  • Description

    Alexa Fluor® 647 Rabbit monoclonal [EP4863(2)] to Melanoma gp100

  • Host species

    Rabbit

  • Conjugation

    Alexa Fluor® 647. Ex: 652nm, Em: 668nm

  • Tested applications

    Suitable for: Flow Cyt, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse

  • Immunogen

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

  • Positive control

    • ICC/IF: B16F10 cells. Flow Cyt: B16F10 cells.

  • General notes

    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.

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

    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 ab246730 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 1/5000.
ICC/IF 1/100.

This product gave a positive signal in B16F10 fixed with 100% methanol (5 mins).

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

    • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Melanoma gp100 antibody [EP4863(2)] (ab246730)
      Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Melanoma gp100 antibody [EP4863(2)] (ab246730)

      ab246730 staining Melanoma gp100 in B16F10 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab246730 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in ). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Flow Cytometry - Alexa Fluor® 647 Anti-Melanoma gp100 antibody [EP4863(2)] (ab246730)
      Flow Cytometry - Alexa Fluor® 647 Anti-Melanoma gp100 antibody [EP4863(2)] (ab246730)

      Overlay histogram showing B16F10 cells stained with ab246730 (red line). The cells were fixed with 80 % methanol (5 min) and then permeabilized with 0.1 % PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10 % normal goat serum to block non-specific protein-protein interaction  followed by the antibody (ab246730) (1x 106 in 100µl at 0.1 µg/ml (1/5000)) for 30 min at 22°C.

      Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor®647 (ab199093) used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.

      Acquisition of >5,000 events were collected using a 40 mW Red laser (640nm) and 670/14 bandpass filter.

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab246730? Please let us know so that we can cite the reference in this datasheet.

    ab246730 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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

    There are currently no Customer reviews or Questions for ab246730.
    Please use the links above to contact us or submit feedback about this product.

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