Overview

  • Product name

    Anti-MAGE3 antibody [EPR20658]
    See all MAGE3 primary antibodies
  • Description

    Rabbit monoclonal [EPR20658] to MAGE3
  • Host species

    Rabbit
  • Specificity

    This antibody shows cross reactivity to MAGEA2 and MAGEA6 as well as weak cross reactivity to MAGEA1 and MAGEA12. It does not cross react with MAGEA4 and MAGEA8.
  • Tested applications

    Suitable for: ICC/IF, IP, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human MAGE3 aa 50 to the C-terminus. The exact sequence is proprietary.
    Database link: P43357

  • Positive control

    • WB: RPMI-8226 and A-375 whole cell lysates; Human MAGE3, MAGEA6 and MAGEA2 recombinant proteins. ICC/IF: HeLa and A-375 cells. IP: A-375 whole cell lysate.
  • General notes

     This product was previously labelled as MAGEA3

     

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 0.05% BSA, 40% Glycerol
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20658
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab213722 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
ICC/IF 1/100.
IP 1/30.
WB 1/1000. Detects a band of approximately 45-50 kDa (predicted molecular weight: 34 kDa).

Target

  • Function

    Proposed to enhance ubiquitin ligase activity of RING-type zinc finger-containing E3 ubiquitin-protein ligases. May enhance ubiquitin ligase activity of TRIM28 and stimulate p53/TP53 ubiquitination by TRIM28. Proposed to act through recruitment and/or stabilization of the Ubl-conjugating enzyme (E2) at the E3:substrate complex. May play a role in embryonal development and tumor transformation or aspects of tumor progression. In vitro promotes cell viability in melanoma cell lines. Antigen recognized on a melanoma by autologous cytolytic T-lymphocytes.
  • Tissue specificity

    Expressed in many tumors of several types, such as melanoma, head and neck squamous cell carcinoma, lung carcinoma and breast carcinoma, but not in normal tissues except for testes and placenta. Never expressed in kidney tumors, Leukemias and lymphomas.
  • Sequence similarities

    Contains 1 MAGE domain.
  • Information by UniProt
  • Database links

  • Alternative names

    • Antigen MZ2 D antibody
    • Antigen MZ2-D antibody
    • Cancer/testis antigen 1.3 antibody
    • Cancer/testis antigen family 1 member 3 antibody
    • CT1.3 antibody
    • HIP8 antibody
    • HYPD antibody
    • MAGA3_HUMAN antibody
    • MAGE 3 antigen antibody
    • MAGE family member A3 antibody
    • MAGE-3 antigen antibody
    • MAGE3 antibody
    • MAGEA3 antibody
    • MAGEA6 antibody
    • Melanoma antigen family A 3 antibody
    • Melanoma associated antigen 3 antibody
    • Melanoma-associated antigen 3 antibody
    • MGC1461 antibody
    see all

Images

  • All lanes : Anti-MAGE3 antibody [EPR20658] (ab213722) at 1/1000 dilution

    Lane 1 : A-375 (human malignant melanoma cell line), whole cell lysate
    Lane 2 : RPMI-8226 (human plasmacytoma, myeloma B lymphocyte), whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 34 kDa
    Observed band size: 45-50 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The molecular mass is consistent with what has been described in the literature (PMID: 17942928).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling MAGE3 with ab213722 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Nuclear and cytoplasmic staining in HeLa cell line (PMID: 23946777, PMID: 23390371) is observed.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • MAGE3 was immunoprecipitated from 0.35 mg of A-375 (human malignant melanoma cell line) whole cell lysate with ab213722 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab213722 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

    Lane 1: A-375 whole cell lysate 10 μg (Input).
    Lane 2: ab213722 IP in A-375 whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab213722 in A-375 whole cell lysate.

    Exposure time: 1 second.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  • All lanes : Anti-MAGE3 antibody [EPR20658] (ab213722) at 1/1000 dilution

    Lane 1 : His-tagged human MAGE3 recombinant protein (aa104-314)
    Lane 2 : His-tagged human MAGEA4 recombinant protein (aa1-317)
    Lane 3 : His-tagged Human MAGEA6 recombinant protein (aa1-314)
    Lane 4 : His-tagged Human MAGEA12 recombinant protein (aa1-314)

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 34 kDa
    Observed band size: 25,39 kDa why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-MAGE3 antibody [EPR20658] (ab213722) at 1/1000 dilution

    Lane 1 : His-tagged human MAGE3 recombinant protein (aa104-314)
    Lane 2 : His-tagged human MAGEA1 recombinant protein (aa1-309)
    Lane 3 : His- and vinc-tagged Human MAGEA2 recombinant protein (aa1-314)
    Lane 4 : His-tagged Human MAGEA8 recombinant protein (aa1-318)

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 34 kDa
    Observed band size: 25,39,75 kDa why is the actual band size different from the predicted?


    Exposure time: 7 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A-375 (human malignant melanoma cell line) cells labeling MAGE3 with ab213722 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Nuclear and cytoplasmic staining in A-375 cell line (PMID: 23946777, PMID: 23390371) is observed.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

References

ab213722 has not yet been referenced specifically in any publications.

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