Anti-CD40 [FGK45]

Flow cytometry using the Anti-CD40 antibody FGK45. Paraformaldehyde fixed mouse splenocytes were stained with anti-unknown specificity antibody (orb256458; isotype control, black line) or the rabbit IgG version of FGK45 (orb1152479, 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.

Immunofluorescence staining of mouse splenocytes with anti-CD40 FGK45. Immunofluorescence analysis of paraformaldehyde fixed mouse splenocytes on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of FGK45 (orb1152479) 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 orb1152479, 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-CD40 antibody FGK45. Mouse lymph node tissue lysate (35 µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of FGK45 (orb1152479) at 2 µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.