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RecombinantRabMAb

Recombinant Anti-MAGE3 antibody [EPR20658] - BSA and Azide free (ab228521)

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Immunocytochemistry/ Immunofluorescence - Anti-MAGE3 antibody [EPR20658] - BSA and Azide free (ab228521)
  • Immunoprecipitation - Anti-MAGE3 antibody [EPR20658] - BSA and Azide free (ab228521)
  • Immunocytochemistry/ Immunofluorescence - Anti-MAGE3 antibody [EPR20658] - BSA and Azide free (ab228521)
  • Anti-MAGE3 antibody [EPR20658] - BSA and Azide free (ab228521)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20658] to MAGE3 - BSA and Azide free
  • Suitable for: ICC/IF, IP, WB
  • Reacts with: Human

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Protein
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Recombinant Human MAGE3 protein (ab99958)

View more associated products

Overview

  • Product name

    Anti-MAGE3 antibody [EPR20658] - BSA and Azide free
    See all MAGE3 primary antibodies
  • Description

    Rabbit monoclonal [EPR20658] to MAGE3 - BSA and Azide free
  • 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

    • ICC/IF: A-375 cells.
  • General notes

    ab228521 is the carrier-free version of ab213722 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab228521 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product was previously labelled as MAGEA3

    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.

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20658
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Cell Type Markers
    • Tumor Associated
    • Cancer
    • Tumor immunology
    • Tumor-associated antigens

Associated products

  • Alternative Versions

    • Anti-MAGE3 antibody [EPR20658] (ab213722)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • FITC Conjugation Kit (Fast) - Lightning-Link® (ab188285)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human MAGE3 protein (ab99958)
  • Related Products

    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
    • Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

Applications

Our Abpromise guarantee covers the use of ab228521 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 Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 45-50 kDa (predicted molecular weight: 35 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.
  • Target information above from: UniProt accession P43357 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4102 Human
    • Omim: 300174 Human
    • SwissProt: P43357 Human
    • Unigene: 417816 Human
    • 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

    • Immunocytochemistry/ Immunofluorescence - Anti-MAGE3 antibody [EPR20658] - BSA and Azide free (ab228521)
      Immunocytochemistry/ Immunofluorescence - Anti-MAGE3 antibody [EPR20658] - BSA and Azide free (ab228521)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213722).

    • Immunoprecipitation - Anti-MAGE3 antibody [EPR20658] - BSA and Azide free (ab228521)
      Immunoprecipitation - Anti-MAGE3 antibody [EPR20658] - BSA and Azide free (ab228521)

      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 1/1000 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213722).

    • Immunocytochemistry/ Immunofluorescence - Anti-MAGE3 antibody [EPR20658] - BSA and Azide free (ab228521)
      Immunocytochemistry/ Immunofluorescence - Anti-MAGE3 antibody [EPR20658] - BSA and Azide free (ab228521)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213722).

    • Anti-MAGE3 antibody [EPR20658] - BSA and Azide free (ab228521)
      Anti-MAGE3 antibody [EPR20658] - BSA and Azide free (ab228521)

    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

    Datasheets and documents

    • Datasheet

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

  • References (0)

    Publishing research using ab228521? Please let us know so that we can cite the reference in this datasheet.

    ab228521 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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