Anti-Prion [3F4]

Immunofluorescence staining of fixed U251 cells with anti-Prion antibody 3F4. Immunofluorescence analysis of paraformaldehyde fixed U251 cells on Shi-fix™ coverslips, permeabilized with 0.15% Triton and stained with the chimeric mouse IgG1 version of 3F4 (orb613863) 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 orb613863, DAPI, merged channels and an isotype control. The isotype control was stained with an unknown specificity antibody (orb256449) followed by Alexa Fluor® 488 secondary antibody.

Western Blot using anti-major Prion protein antibody 3F4 Human brain cerebral cortex (A), cerebellum (B), and hippocampus (C) lysate samples (35 µg protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric mouse IgG1 version of 3F4 (orb613863). Cerebral cortex and hippocampus samples were probed using 0.001 µg/ml of orb613863, and cerebellum samples with 0.003 µg/ml of orb613863, before detection using an anti-mouse secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence. The predicted running size for unmodified major Prion protein is 27.7 kDa though this protein has several glycosylated forms, may be lipidated, sumoylated and post-translationally cleaved to produce the matrue form of the protein [Uniprot]. orb613863 successfully detected major Prion protein in human brain cerebral cortex, cerebellum, and hippocampus lysates.