Antibodies, Proteins, Kits and Reagents for Life Science

278 083 ₸ Product size

100 µl 278 083 ₸

Order now and get it on Tuesday March 09, 2021

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

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 & Species

Application	Species
ICC/IF	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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

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.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20658
Isotype

IgG
Research areas

Western blot - Anti-MAGE3 antibody [EPR20658] (ab213722)

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).
Immunocytochemistry/ Immunofluorescence - Anti-MAGE3 antibody [EPR20658] (ab213722)

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.
Immunoprecipitation - Anti-MAGE3 antibody [EPR20658] (ab213722)

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.
Western blot - Anti-MAGE3 antibody [EPR20658] (ab213722)

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.
Western blot - Anti-MAGE3 antibody [EPR20658] (ab213722)

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.
Immunocytochemistry/ Immunofluorescence - Anti-MAGE3 antibody [EPR20658] (ab213722)

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.
Anti-MAGE3 antibody [EPR20658] (ab213722)

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