United Kingdom (UK)
Recombinant monoclonal antibody to CD3. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from CD3B219.
UniProt Accession Number of Target Protein: P07766
Alternative Name(s) of Target: T-cell surface glycoprotein CD3 epsilon chain; T-cell surface antigen T3/Leu-4 epsilon chain; CD3e; SP34-2
Specificity: The antibody is specific for CD3 epsilon.
Application Notes: The antibody was used to construct different CD33 xCD3 bispecific antibodies (IgG4). In vitro T cell mediated cytotoxicity assays were performed using the bispecific antibody and the cytotoxicity was measured by flow cytometry. Ex vivo cytotoxicity assay showed the paired antibody promoted a dose-dependent reduction of total cytotoxicity that correlated with T cell activation after 48 hours (US20210040209A1). The antibody is considered a high affinity binder (Kd= 8mM) (US20210214440A1). The CD33×CD3 bispecific antibodies specifically induce cell cytotoxicity of MOLM-13 cells and CD33+ monocytes at 48 hr (US20200048349A1).
Antibody first published in: PMID:
Flow cytometry using anti-CD3 antibody CD3B219 (Ab03166). Jurkat cells were stained with the anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of CD3B219 (Ab03166-23.0, 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 the cells were analyzed using a FACSCanto flow cytometer.
Western blot using anti-CD3 antibody CD3B219 (Ab03166). Jurkat (A) (0.001µg/ml) and Molt-4 (B) (0.001µg/ml) cells lysate, and human lymph node (C) (0.1µg/ml) tissue lysate (35µg protein in RIPA buffer) were resolved on an SDS-PAGE gel, and blots were probed with the chimeric rabbit version of CD3B219 (Ab03166-23.0) at the mentioned respective concentrations before detection using an anti-rabbit secondary antibody. A primary incubation of 1 hour was used, and protein was detected by chemiluminescence.
Immunofluorescence staining of Jurkat cells with anti-CD3 antibody CD3B219 (Ab03166). Immunofluorescence analysis of paraformaldehyde fixed Jurkat cells on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of CD3B219 (Ab03166-23.0) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing membrane staining. The nuclear stain is DAPI (blue). Panels show, from left-right, top-bottom, Ab03166-23.0, DAPI, merged channels, and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.