The C-terminal Myc-tagged Recombinant, derived from the c-Myc protein, is a popular epitope tag for detecting recombinant protein expression in yeast, bacterial, insect, and mammalian cell systems. The Myc tag can be fused to the N-terminus or the C-terminus of a protein. The well-characterized Myc tag provides a reliable method for the detection and purification of tagged target proteins without a protein-specific antibody or probe. Our Myc-tagged antibodies are designed to reliably detect recombinant Myc-tagged proteins. Each antibody is validated for use in various applications.
Immunofluorescent analysis of c-Myc (green) in HEL 11.4-induced IPS cells cultured for a few days on Matrigel-coated chamber slides.
Cells fixed in 4% paraformaldehyde were permeabilized with 0.1% Triton X-100 for 15 minutes at room temperature. Cells were probed with a monoclonal antibody c-Myc (9E10) (cat. no. MA1-980) at a dilution of 1:200 overnight at 4°C, washed with PBST and incubated with an antibody fluorescein-conjugated secondary at a dilution of 1:100 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI and cells were analyzed by fluorescence microscopy at 20X magnification.
Western blot analysis of c-Myc.
Performed by loading 25 µg of 293t cells transfected with c-Myc-MYCBP (lane 1), transfected with c-Myc-FLI1 (lane 2) and transfected with 12Tag (lane 3) on an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with a polyclonal Myc Tag antibody (Cat# PA1-981) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hour at room temperature. The temperature is in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western blot substrate (Cat. No. 32132). The image shows target bands at c-Myc–MYCBP (19kDa), c-Myc-FLI1 (54kDa), and 12Tag (45kDa), which are consistent with the predicted molecular weights for each target.
Flow cytometric analysis of c-Myc (blue histogram) in H9 embryonic stem cells.
To generate single-cell suspensions, colonies were treated with TrypLE cell-dissociating enzyme for 5 minutes at 37°C. Cells were incubated with a monoclonal antibody c-Myc(9E10) (Cat. No. MA1-980) or mouse IgG (green histogram) at a dilution of 1:100 for 1 hour on ice, washed with PBS + 5 % calf fetus. serum (FACS buffer) and incubated with a fluorescein-conjugated secondary antibody at a dilution of 1:200 for 30 minutes on ice. Cells were washed with cold FACS buffer, resuspended in 500 µl FACS buffer containing 10 µl 4% paraformaldehyde and analyzed in a flow cytometer.