Recombinant Anti-MAGE3 antibody [EPR20658] (ab213722)
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
- Suitable for: ICC/IF, IP, WB
- Reacts with: Human
Related conjugates and formulations
Overview
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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. This information is proprietary to Abcam and/or its suppliers.
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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.
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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
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
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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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20658 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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 |
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ICC/IF |
1/100.
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IP |
1/30.
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WB |
1/1000. Detects a band of approximately 45-50 kDa (predicted molecular weight: 34 kDa).
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Notes |
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ICC/IF
1/100. |
IP
1/30. |
WB
1/1000. Detects a band of approximately 45-50 kDa (predicted molecular weight: 34 kDa). |
Target
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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
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Database links
- Entrez Gene: 4102 Human
- Omim: 300174 Human
- SwissProt: P43357 Human
- Unigene: 417816 Human
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Alternative names
- Antigen MZ2 D antibody
- Antigen MZ2-D antibody
- Cancer/testis antigen 1.3 antibody
see all
Images
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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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The molecular mass is consistent with what has been described in the literature (PMID: 17942928).
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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.
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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.
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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 secondBlocking/Dilution buffer: 5% NFDM/TBST.
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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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.
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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.
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab213722 has not yet been referenced specifically in any publications.