Anti-CD3 [CD3B219]

Flow cytometry using anti-CD3 antibody CD3B219. Jurkat cells were stained with the anti-unknown specificity antibody (orb256458; isotype control, black line) or the rabbit IgG version of CD3B219 (orb1821575, 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.

Immunofluorescence staining of Jurkat cells with anti-CD3 antibody CD3B219. Immunofluorescence analysis of paraformaldehyde fixed Jurkat cells on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of CD3B219 (orb1821575) (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, orb1821575, DAPI, merged channels, and an isotype control. The isotype control was an unknown specificity antibody (orb256458) followed by staining with Alexa Fluor® 488 secondary antibody.

Western blot using anti-CD3 antibody CD3B219. Jurkat (A) (0.001 µg/ml) and Molt-4 (B) (0.001 µg/ml) cells lysate, and human lymph node (C) (0.1 µg/ml) tissue lysate (35 µg protein in RIPA buffer) were resolved on an SDS-PAGE gel, and blots were probed with the chimeric rabbit version of CD3B219 (orb1821575) 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.