A myc tag is a polypeptide protein tag derived from the c-myc gene product that can be added to a protein using recombinant DNA technology. It can be used for affinity chromatography, then used to separate recombinant, overexpressed protein from wild type protein expressed by the host organism. It can also be used in the isolation of protein complexes with multiple subunits. A myc tag can be used in many different assays that require recognition by an antibody. If there is no antibody against the studied protein, adding a myc-tag allows one to follow the protein with an antibody against the Myc epitope. Examples are cellulite localization studies by immunofluorescence or detection by Western blotting. The peptide sequence of the myc-tag is (in 1- and 3-letter codes, respectively): EQKLISEEDL and Glu-Gln-Lys-Leu-Ile-Ser-Glu-Glu-Asp-Leu. The tag is approximately 1202 Daltons in atomic mass and has 10 amino acids. It can be fused to the C-terminus and the N-terminus of a protein. It is advisable not to fuse the tag directly behind the signal peptide of a secretory protein, since it can interfere with translocation into the secretory pathway. A monoclonal antibody against the myc epitope, named 9E10, is available from the non-commercial Developmental Studies Hybridoma Bank.
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别名
c-myc tag antibody
Myc Epitope Tag antibody
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Western blot analysis of Myc tag on different lysates with Mouse anti-Myc tag antibody (HA601513) at 1/20,000 dilution.
Lane 1: 293T transfected with Myc-tagged ACAT2 (C-terminal) cell lysate Lane 2: 293T transfected with Myc-tagged Claudin18.2 (C-terminal) cell lysate Lane 3: 293T transfected with Myc-tagged Histone H3.1 (N-terminal) cell lysate
Lysates/proteins at 10 µg/Lane.
Exposure time: 25 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601513) at 1/20,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of HeLa cells labeling Myc tag with Mouse anti-Myc tag antibody (HA601513) at 1/200 dilution.
HeLa cells, transfected with empty control (top, negative) / Myc-tagged Claudin18.2 (C-terminal) (middle, positive) / Myc-tagged Histone H3.1 (N-terminal) (bottom, positive) expression vector, respectively, were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-Myc tag antibody (HA601513) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Myc tag was immunoprecipitated from 0.2 mg 293T transfected with Myc-tagged Histone H3.1 (N-terminal) cell lysate with HA601513 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA601513 at 1/20,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: 293T transfected with Myc-tagged Histone H3.1 (N-terminal) cell lysate (input) Lane 2: HA601513 IP in 293T transfected with Myc-tagged Histone H3.1 (N-terminal) cell lysate Lane 3: Mouse IgG instead of HA601513 in 293T transfected with Myc-tagged Histone H3.1 (N-terminal) cell lysate