Sensitive FLAG Antibody For Blotting Needs

The product reviewed here is an anti-FLAG antibody that recognizes DYKDDDDK tag either at the N- or C-terminus of target proteins. This antibody was tested in protein detection from lysates resolved by SDS-PAGE and transferred on to nitrocellulose membrane. The LiCOR Odyssey scanner was used for detection of the FLAG-tagged protein. Overall, the antibody performed very well in detecting FLAG-tagged Cas9 expressed transiently in 293T cells. Four different clones were tested and detection was compared with an antibody from a different vendor. In summary, the BioLegend FLAG L5 antibody was found to be much more sensitive than the other antibody tested in the same experiment. This antibody was also used in immunofluorescence. A FLAG-tagged ER-marker protein was transiently expressed in HeLa cells cultured on glass coverslips and the cells were fixed with 4% paraformaldehyde. FLAG tag was recognized using the anti-FLAG-L5 antibody (1:500 dilution) and a Calnexin antibody was used as a control for ER staining. Alexa-tagged secondary antibodies (1:1000 dilution) were used to fluorescently label the primary antibodies bound to respective antigens. Images were acquired using an epifluorescence microscope.