Anti-RAP [7F1]

Flow cytometry using the anti-RAP antibody 7F1. HeLa cells were fixed using 2% PFA and stained with anti-unknown specificity antibody (orb256458; isotype control, black line) or the rabbit IgG1 version of 7F1 (orb758871, 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 fixed HeLa cells with anti-RAP antibody 7F1. Immunofluorescence analysis of paraformaldehyde fixed HeLa cells on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of 7F1 (orb758871) 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 orb758871, 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-RAP antibody Staining in HeLa(A) (0.1 ug/ml), A549 (B) (0.01 ug/ml), HepG2 (C) (0.01 ug/ml), HEK293 (D) (0.03 ug/ml), MCF7 (E) (0.03 ug/ml), Caco-2 (F) (0.001 ug/ml) and U251 (G) (0.001 ug/ml) cell lysates (35 µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of RAP (orb758871) at 0.5 µg/ml, before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.