United Kingdom (UK)
Recombinant monoclonal antibody to CD131. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma BION-1.
UniProt Accession Number of Target Protein: P32927
Alternative Name(s) of Target: CSF2RB; CD131; IL3RB; IL-3R common beta subunit; IL5RB; colony stimulating factor 2 receptor, beta, low-affinity (granulocyte-macrophage); SMDP5; Common Beta-Chain; CDw131; cytokine receptor common subunit beta; beta-c; CD131; Beta Common Chain
Immunogen: BALB/c mice were immunized intraperitonally with COS cells transfected DQP (Domain 4 of the common beta-chain) expression constructs. The immunizations were repeated 4 times at 2-week intervals. Four weeks after the final immunization, a mouse was boosted with COS transfectants intravenously. Three days later, splenocytes were harvested and fused with NS-1 myeloma cells. Hybridoma supernatants were screened on CHO DQP cells by flow cytometry, with untransfected CHO cells as a control. All antibodies were from single hybridoma clones as selected by a limiting dilution method. MoAbs were purified from ascites fluid or hybridoma supernatant by a protein A sepharose column.
Specificity: Binds to domain 4 of the common beta-chain and recognises the same binding site used by IL-5, GM-CSF and IL-3, or binds to an epitope in close proximity to it. Residues M363, R364, E366 (B-C loop) and R418 (F-G loop) in the common beta-chain are important for BION-1 binding.
Application Notes: BION-1 has been shown the block eosinophil production, survival, and activation stimulated by IL-5 as well as by GM-CSF and IL-3, thereby influencing inflammation (Sun et al., 1999; PMID: 10477723). Further, BION-1 has been used in immunohistochemistry experiments in order to examine the presence of GM-CSF-receptor protein in human 8-cell embryos and blastocysts (Sjoblom et al., 2002; PMID: 12444058). In addition, a fab fragment derived from BION-1 has been used in the crystallisation of the common beta-chain to analyse the structure of the activation domain of the GM-CSF/IL-3/IL-5 receptor (Rossjohn et al., 2000; PMID: 10753826).
Antibody first published in:
Sun et al. Simultaneous Antagonism of Interleukin-5, Granulocyte-Macrophage Colony-Stimulating Factor, and Interleukin-3 Stimulation of Human Eosinophils by Targetting the Common Cytokine Binding Site of Their Receptors Blood, Vol 94, No 6 (September 15), 1999: pp 1943-1951 PMID:10477723
Note on publication:
Describes the presence of the common beta-chain in various cell surface receptors and its role in receptor activation for inflammation. The use of BION-1 as an antagonist of IL-5, GM-CSF and IL-3 signalling through binding the common beta-chain is also outlined in the context of blocking eosinophil activation.
Immunofluorescence staining of U937 cells using anti-CD131 antibody (Ab00957) BION-1. Immunofluorescence analysis of paraformaldehyde fixed U937 cells immobilized on Shi-fix™ cover-slips and stained with the chimeric rabbit IgG version of BION-1 (Ab00957-23.0) at 10 µg/ml followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab00957-23.0, 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-CD131 antibody BION-1 (Ab00957). U937 cells were stained with anti-Fluorescein IgG antibody (4-4-20; isotype control, black line) or the rabbit IgG version of BION-1 ((Ab00957-23.0, 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.