Anti-Melanoma gp100 antibody [EP4863(2)] - Low endotoxin, Azide free (ab193201)

Rabbit monoclonal Melanoma gp100 antibody [EP4863(2)]. Validated in WB, IP, IHC, Flow Cyt, ICC/IF and tested in Mouse, Human.

Overview

  • Product name

    Anti-Melanoma gp100 antibody [EP4863(2)] - Low endotoxin, Azide free
    See all Melanoma gp100 primary antibodies
  • Description

    Rabbit monoclonal [EP4863(2)] to Melanoma gp100 - Low endotoxin, Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, ICC/IF, Flow Cyt, IP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

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

  • Positive control

    • B16-F0 and Human melanoma lysates; Human melanoma tissue.
  • General notes

    ab193201 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

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

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

Properties

Applications

Our Abpromise guarantee covers the use of ab193201 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 Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. 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

  • This IHC data was generated using the same anti-Melanoma gp100 antibody clone, EP4863(2), in a different buffer formulation (cat# ab137078).

    Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human melanoma tissue labelling Melanoma gp100 with ab137078 at 1/100 dilution.

  • ab137078 staining Melanoma gp100 in B16-F0 (mouse skin) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. ab7291 and ab150120 were used as counterstains for primary antibody ab137078 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

    Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
    Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137078).

  • ab137078 staining Melanoma gp100 in the mouse cell line B16-F0 (mouse skin) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/80. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/500 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137078).

  • ab137078 immunoprecipitating Melanoma gp100. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.

    Lane 1: B16-F0 (mouse skin) whole cell lysate 10ug
    Lane 2: B16-F0 (mouse skin) whole cell lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab137078 in B16-F0 whole cell lysate

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137078).

  • Overlay histogram showing B16-F0 (Mouse skin melanoma cell) cells stained with ab137078 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with ab137078 at 1/450 dilution. The secondary antibody used was Goat anti rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was Rabbit monoclonal IgG (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137078).

  • Overlay histogram showing MALME 3M cells stained with ab137078 (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 followed by the antibody (ab137078, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137078).

  • This ICC/IF data was generated using the same anti-Melanoma gp100 antibody clone, EP4863(2), in a different buffer formulation (cat# ab137078).

    ab137078 staining Melanoma gp100 in B16-F0 (mouse skin) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. ab7291 and ab150120 were used as counterstains for primary antibody ab137078 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

    Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
    Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

References

ab193201 has not yet been referenced specifically in any publications.

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