United Kingdom (UK)
Recombinant monoclonal antibody to MAGEA3. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the human B cell clone 21B4.
UniProt Accession Number of Target Protein: P43357; P43360
Alternative Name(s) of Target: MAGE-3; MAGE3; MAGE 3; CT1.3; MAGE-3 antigen; Melanoma-associated antigen 3; Antigen MZ2-D; Cancer/testis antigen 1.3; MAGEA6; MAGE6; Melanoma-associated antigen 6; Cancer/testis antigen 1.6; CT1.6; MAGE-6 antigen; MAGE3B antigen
Immunogen: The original antibody was isolated from a patient suffering from a tumor expressing a tumor specific antigen like MAGE-A3.
Specificity: This antibody binds human MAGEA3 and MAGEA6, but does not cross react with other members of the MAGE family like MAGE-A1, MAGE-A4 and MAGE-A10. MAGE-A3 is a tumor specific antigen and it is expressed on many tumors like melanoma, non-small cell lung cancer, hematologic malignancies, and others.
Application Notes: The binding specificity of this antibody towards MAGE-A3 was determined using ELISA. The original antibody cross reacts with MAGEA6 but does not react with other members of the MAGE family in an ELISA or western blot assay. This antibody was used in the detection of HEK293T cells expressing MAGEA3 and MAGEA6 using flow cytometry. The original IgG1 antibody was capable of immunoprecipitating MAGE-A3 from lysates of SK-MEL-37 cells endogenously expressing MAGEA3. The original antibody was also used in the immunohistochemical staining of tissues form normal testis and squamous cell carcinoma of the lung (US20140093514).
Antibody first published in: Esslinger et al. Magea3 binding antibodies. (US20140093514) PMID:
Western Blot using anti-MAGE-3 antibody 21B4 (Ab02838). HeLa(A) (0.01µg/ml) cells lysate and HeLa(B) (0.01µg/ml) cells nuclear lysate (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of 21B4 (Ab02838-23.0) before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Immunofluorescence staining of HeLa cells with anti-MAGE-3 (Ab02838) 21B4 Immunofluorescence analysis of paraformaldehyde fixed HeLa cells stained with the chimeric rabbit IgG version of 21B4 (Ab02838-23.0) (1:1000 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1500 dilution), showing nuclear staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab02838-23.0, DAPI, merged channels and an isotype control. The isotype control was anti-fluorescein antibody (Ab00102-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.
Flow-cytometry using the anti-MAGE-3 antibody 21B4 (Ab02838). HeLa cells were stained with an isotype control (Ab00102-23.0, black line) or the rabbit-chimeric version of 21B4 (Ab02838-23.0, blue line) at a concentration of 1 µg/ml for 30 mins at RT. After washing, bound antibody was detected using Alexa Fluor 488® conjugated goat anti-rabbit IgG and cells analysed on a FACSCanto flow-cytometer.