United Kingdom (UK)
Recombinant monoclonal antibody to Mouse antibody kappa light chain. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma OX-20.
UniProt Accession Number of Target Protein: P01837
Alternative Name(s) of Target: kappa Lc; OX20; MRC OX-20; MRC OX20
Immunogen: OX-20 was generated by hyperimmunizing rats with mouse F(ab')2 generated by enzymatic digest.
Specificity: OX-20 binds specifically to mouse kappa light chain of immunoglobulins. Kappa light chains are found in 95% of mouse Igs with lambda light chains being found in other antibodies. Kappa and lambda chains differ slightly in amino acids residue composition.
Application Notes: OX-20 can be used for WB, IP and FC studies.
Antibody first published in:
Mason et al. The rat mixed lymphocyte reaction: roles of a dendritic cell in intestinal lymph and T-cell subsets defined by monoclonal antibodies. Immunology 44:75-87 1981 PMID:6456222
Note on publication:
Describes how rat anti-mouse antibodies were shown to bind specifically to IgGs and not to IgM by indirect haemagglutination of sheep red blood cells which had been coated with different mouse mAbs specific for sheep red blood cells, and to mouse IgGs and not IgG molecules derived from other species (some cross reaction with rabbit and guinea pig) (Galfre et al. 1979).
Western Blot using anti-Mouse antibody kappa light chain antibody OX-20 (Ab00583) Mouse spleen (A) and mouse thymus (B) tissue lysates (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of OX-20 (Ab00583-23.0) at 0.001 µg/ml and 0.0001 µg/ml, respectively, before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Immunofluorescence staining of fixed mouse splenocytes with anti-Mouse antibody kappa light chain antibody OX-20 (Ab00583) Immunofluorescence analysis of paraformaldehyde fixed mouse splenocytes on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of OX-20 (Ab00583-23.0) at 10 µg/ml for 1h 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 Ab00583-23.0, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-1.1) followed by staining with Alexa Fluor® 488 secondary antibody.
Flow cytometry using the Anti-Mouse antibody kappa light chain antibody OX-20 (Ab00583). mouse splenocytes were stained with anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of OX-20 (Ab00583-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, the 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.
ELISA using OX-20 (Ab00583), Ab00102-1.1, Ab00102-2.0 and Ab00104-1.1. Binding curves of the rabbit IgG chimeric version of the Anti-HA antibody OX-20 against Ab00102-1.1 (blue line), Ab00102-2.0 (orange line) and Ab00104-1.1 (grey line) to an ELISA plate coated with Ab00102-1.1, Ab00102-2.0 and Ab00104-1.1 at a concentration of 2.5 ug/ml. A 3-fold serial dilution from 3000 to 0.0169 ng/ml was performed using Ab00583 antibody. For signal detection, a 1:4000 dilution of a HRP-conjugated anti-rabbit IgG antibody was used.