United Kingdom (UK)
Recombinant monoclonal antibody to RAGE. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma 2A11.
UniProt Accession Number of Target Protein: Q15109
Alternative Name(s) of Target: AGER; advanced glycosylation end product-specific receptor; receptor for advanced glycosylation end products
Immunogen: Human RAGE protein.
Specificity: 2A11 is specific for the V-domain of human advanced glycosylation end product-specific receptor. It recognises both the glycosylated and non-glycosylated forms.
Application Notes: ELISA confirmed the specificity of 2A11 for the V-domain of RAGE, with a binding affinity (Kd) of 7.6 nM. ELISA was also used to confirm that the binding of 2A11 to RAGE is glycosylation independent. In SPR experiments the Kd of 2A11 binding to RAGE was determined as 0.38 nM. Flow cytometry and immunochemistry were used to visualise the binding of 2A11 to HEK-RAGE cells.
Antibody first published in:
Jyoti, 2012. Development of new antibody based theranostic agents targeting the receptor for advanced glycation end-product (RAGE). Dissertation Submitted to the Graduate Faculty of the North Dakota State University of Agriculture and Applied Science PMID:
Note on publication:
Describes the characterisation of 2A11 through ELISA, SPR, FC, and IH experiments.
Western Blot using anti-RAGE antibody 2A11 (Ab01230). Human lung lysate (35µg protein in RIPA buffer) was resolved on an SDS PAGE gel and blots probed with the chimeric rabbit IgG version of 2A11 (Ab01230-23.0) at 0.03 µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Immunofluorescence staining of A549 cells using anti-RAGE antibody (Ab01230) 2A11. Immunofluorescence analysis of paraformaldehyde fixed A549 cells stained with the chimeric rabbit IgG version of 2A11 (Ab01230-23.0) at 10 µg/ml followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing cytoplasmic staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab01230-23.0, DAPI, merged channels and an isotype control. The isotype control was stained with anti-Fluorescein antibody (Ab00102-23.0) followed by Alexa Fluor® 488 secondary antibody.
Flow-cytometry using the Anti-RAGE antibody 2A11 (Ab01230). A549 cells were stained with anti-Fluorescein IgG antibody (4-4-20; isotype control, shaded line) or the rabbit IgG version of 2A11 (Ab01230-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.