The Myc tag, derived from the c-Myc protein, is a popular epitope tag for detecting the expression of recombinant proteins in yeast, bacteria, insect, and mammalian cell systems. The Myc tag may be fused to either the N-terminus or 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 tag antibodies are designed to dependably detect recombinant Myc-tagged proteins. Each antibody is validated for use in various applications.
Invitrogen Myc tag antibodies are designed to dependably detect recombinant Myc-tagged proteins in various applications.
Immunofluorescent analysis of c-Myc (green) in HEL 11.4 induced IPS cells grown 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 c-Myc Monoclonal Antibody (9E10) (Cat. No. MA1-980) at a dilution of 1:200 overnight at 4°C, washed with PBST, and incubated with a fluorescein-conjugated secondary antibody 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) onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with a Myc Tag Polyclonal Antibody (Cat. No. 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 in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Cat. No. 32132). The image shows the target bands on c-Myc –MYCBP (19kDa), c-Myc -FLI1 (54kDa) and 12Tag (45kDa), which are consistent with predicted molecular weights for each target.
Flow cytometric analysis of c-Myc (blue histogram) on H9 embryonic stem cells. To generate single cells suspensions, colonies were treated with TrypLE cell dissociation enzyme for 5 minutes at 37°C. Cells were incubated with a c-Myc Monoclonal Antibody (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% fetal calf 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 of FACS buffer containing 10 µL of 4% paraformaldehyde, and analyzed on a flow cytometer.