Recombinant Human Melanoma gp100 protein (ab132146)

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SDS-PAGE - Recombinant Human Melanoma gp100 protein (ab132146)

    Key features and details

    • Expression system: Wheat germ
    • Tags: GST tag N-Terminus
    • Suitable for: SDS-PAGE, ELISA, WB

    Description

    • Product name

      Recombinant Human Melanoma gp100 protein

    • Expression system

      Wheat germ

    • Accession

      P40967

    • Protein length

      Full length protein

    • Animal free

      No

    • Nature

      Recombinant

      • Species

        Human

      • Sequence

        MDLVLKRCLLHLAVIGALLAVGATKVPRNQDWLGVSRQLRTKAWNRQLYP EWTEAQRLDCWRGGQVSLKVSNDGPTLIGANASFSIALNFPGSQKVLPDG QVIWVNNTIINGSQVWGGQPVYPQETDDACIFPDGGPCPSGSWSQKRSFV YVWKTWGQYWQVLGGPVSGLSIGTGRAMLGTHTMEVTVYHRRGSRSYVPL AHSSSAFTITDQVPFSVSVSQLRALDGGNKHFLRNQPLTFALQLHDPSGY LAEADLSYTWDFGDSSGTLISRALVVTHTYLEPGPVTAQVVLQAAIPLTS CGSSPVPGTTDGHRPTAEAPNTTAGQVPTTEVVGTTPGQAPTAEPSGTTS VQVPTTEVISTAPVQMPTAESTGMTPEKVPVSEVMGTTLAEMSTPEATGM TPAEVSIVVLSGTTAAQVTTTEWVETTARELPIPEPEGPDASSIMSTESI TGSLGPLLDGTATLRLVKRQVPLDCVLYRYGSFSVTLDIVQGIESAEILQ AVPSGEGDAFELTVSCQGGLPKEACMEISSPGCQPPAQRLCQPVLPSPAC QLVLHQILKGGSGTYCLNVSLADTNSLAVVSTQLIMPGQEAGLGQVPLIV GILLVLMAVVLASLIYRRRLMKQDFSVPQLPHSSSHWLRLPRIFCSCPIG ENSPLLSGQQV

      • Predicted molecular weight

        97 kDa including tags

      • Amino acids

        1 to 661

      • Tags

        GST tag N-Terminus

    Specifications

    Our Abpromise guarantee covers the use of ab132146 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    • Applications

      SDS-PAGE

      ELISA

      Western blot

    • Form

      Liquid
    • Concentration information loading...

    Preparation and Storage

    • Stability and Storage

      Shipped on dry ice. Upon delivery aliquot and store at -80ºC. Avoid freeze / thaw cycles.

      pH: 8.00
      Constituents: 0.31% Glutathione, 0.79% Tris HCl

    General Info

    • Alternative names

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

    • 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

    Images

    • SDS-PAGE - Recombinant Human Melanoma gp100 protein (ab132146)
      SDS-PAGE - Recombinant Human Melanoma gp100 protein (ab132146)
      12.5% SDS-PAGE analysis of ab132146 stained with Coomassie Blue.

    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 (1)

    Publishing research using ab132146? Please let us know so that we can cite the reference in this datasheet.

    ab132146 has been referenced in 1 publication.

    • Fässler M  et al. Antibodies as biomarker candidates for response and survival to checkpoint inhibitors in melanoma patients. J Immunother Cancer 7:50 (2019). PubMed: 30786924

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Question

    I wish to do studies of cross-presentation by B cells after stimulation with full length proteins.

    For those studies I am considering your products:

    Melanoma Gp100 (ab38124 + ab132146) and Mart-1 (ab 114312).

    Now I wondered whether these products would be suited for stimulation assays (after which CD8+ T-cell responses are analysed).

    If yes - which is better - tagged or not tagged??

    Read More
    Answer

    Please note that ab38124 is not a full length protein but a recombinant fragment, corresponding to amino acids 1-150 of Human Melanoma gp100.

    As for the stimulation assays, unfortunately we have not tested these proteins in stimulation assays and therefore cannot comment on whether they would be suitable. These proteins have only been validated and guaranteed to work in western blot and ELISA although ab38124 as been validated in an inhibition assay.

    Read More

    Question

    Inquiry: Dear sir or madam, I contact you because I am looking for a method to assay the gp 100 peptide and a CpG oligonucleotide. I should prepare liposomes loaded with this peptide, and I'm not sure how to quantify these products. I was thinking about using ELISA to quantify them, but i do not know which kind of antibody should I use. Could you help me, please? Yours sincerely.

    Read More
    Answer

    Thank you for your inquiry.

    I can confirm that it is possible to assay proteins via ELISA.

    We have the following products that will be suitable for gp100:

    ab52058 (Anti-Melanoma gp100 antibody) is tested and guaranteed for ELISA and human gp100.

    https://www.abcam.com/Melanoma-gp100-antibody-ab52058.html (or use the following: https://www.abcam.com/Melanoma-gp100-antibody-ab52058.html).

    In order to perform an ELISA the liposomes loaded with gp100 will have to be lysed and then the plates coated with the protein. I can also strongly suiggest to use a standard/positive control with a known concentration for this.

    The following ab132146 (Melanoma gp100 protein (Tagged)) protein is suitable as standard:

    https://www.abcam.com/Recombinant-Human-Melanoma-gp100-protein-ab132146.html (or use the following: https://www.abcam.com/Recombinant-Human-Melanoma-gp100-protein-ab132146.html).

    Protocols can be found here:

    https://www.abcam.com/index.html?pageconfig=popular_protocols

    https://www.abcam.com/index.html?pageconfig=resource&rid=12232

    Unfortunately, we do not have antibodies against CpG oligonucleotides. Since gp100 and CpG oligonucleotides are used for immunisations, I am not sure that the antiboduy and protein above it the correct one for you. Please cross check if it is the same gp100!

    I hope this information is helpful and wish you good luck for your research.

    Read More

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