Western Blot using anti-CD34 antibody My10 Jurkat cell extract (35ug protein in RIPA buffer) was resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of My10 at 0.1ug/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence. The expected band size for the canonical isoform of CD34 is 40.9kD, though a second isoform at 35.2kD is also present, with a 56 amino acid truncation at the C-terminal of the protein. CD34 is also glycosylated at several positions. Successfully detected both isoforms of human CD34.

Flow cytometry using anti-CD34 antibody My10 Human leukocytes were stained with an isotype control (panel A) or the rabbit-chimeric version of My10 (panel B) at a concentration of 1ug/ml for 30mins at RT. After washing, bound antibody was detected using a AF488 conjugated donkey anti-rabbit antibody and cells analysed on a FACSCanto flow cytometry.