WB
Western blot analysis of Myc tag on different lysates with Mouse anti-Myc tag antibody at 1/100,000 dilution. Lane 1: 293T transfected with Myc-tagged empty control 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: 1 minute 2 seconds; 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 at 1/100,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of 293T cells labeling Myc tag with Mouse anti-Myc tag antibody at 1/1,000 dilution. 293T cells, transfected with Myc-tagged empty control, Claudin18.2 (C-terminal) or Histone H3.1 (N-terminal) expression vector, respectively, were fixed in 4% paraformaldehyde for 10 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (488) 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 (red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.IP
Myc-tag was immunoprecipitated in 2µg C-terminal Myc-tag fusion protein lysate with Mouse anti-Myc tag antibody. Western blot was performed from the immunoprecipitate using Mouse anti-Myc tag antibody at 1/2,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody at 1:5,000 dilution was used for 60 mins at room temperature. Lane 1: C-terminal Myc-tag fusion protein lysate (input). Lane 2: Mouse anti-Myc tag antibody IP in C-terminal Myc-tag fusion protein lysate. Lane 3: Mouse IgG instead of Mouse anti-Myc tag antibody IP in C-terminal Myc-tag fusion protein lysate. Blocking/Dilution buffer: 5% NFDM/TBST.| Product Name | Myc tag Recombinant Mouse Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide EQKLISEEDL conjugated to keyhole limpet haemocyanin. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG1 |
| Host Species | Mouse |
| Tested Applications | ICC/IFIPWB |
| WB:1:100000-1:200000 ICC/IF:1:1000 IP:Use at an assay dependent concentration. |
|
| Species Reactivity | All |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | c-myc tag antibody Myc Epitope Tag antibody |
|---|---|
| Function | 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. |
WB
Western blot analysis of Myc tag on different lysates with Mouse anti-Myc tag antibody at 1/100,000 dilution. Lane 1: 293T transfected with Myc-tagged empty control 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: 1 minute 2 seconds; 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 at 1/100,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of 293T cells labeling Myc tag with Mouse anti-Myc tag antibody at 1/1,000 dilution. 293T cells, transfected with Myc-tagged empty control, Claudin18.2 (C-terminal) or Histone H3.1 (N-terminal) expression vector, respectively, were fixed in 4% paraformaldehyde for 10 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (488) 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 (red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
IP
Myc-tag was immunoprecipitated in 2µg C-terminal Myc-tag fusion protein lysate with Mouse anti-Myc tag antibody. Western blot was performed from the immunoprecipitate using Mouse anti-Myc tag antibody at 1/2,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody at 1:5,000 dilution was used for 60 mins at room temperature. Lane 1: C-terminal Myc-tag fusion protein lysate (input). Lane 2: Mouse anti-Myc tag antibody IP in C-terminal Myc-tag fusion protein lysate. Lane 3: Mouse IgG instead of Mouse anti-Myc tag antibody IP in C-terminal Myc-tag fusion protein lysate. Blocking/Dilution buffer: 5% NFDM/TBST.| Application Notes | WB:1:100000-1:200000 ICC/IF:1:1000 IP:Use at an assay dependent concentration. |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
