DYKDDDDK Tag ( FLAG ) Recombinant Mouse Monoclonal Antibody [PS00-M2]
cat.: HA601080
| Product Type: | Recombinant Mouse monoclonal IgG1, primary antibodies |
|---|---|
| Species reactivity: | Species independent |
| Applications: | WB, IP, IF-Cell |
| Clonality: | Monoclonal |
| Clone number: | PS00-M2 |
| Form: | Liquid |
| Storage condition: | 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. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Isotype: | IgG1 |
| Immunogen: | Synthetic peptide immune sequence is N-DYKDDDDK-C. |
| Positive control: | C-terminal FLAG-tag fusion protein lysate, N-terminal FLAG-tag fusion protein lysate. |
| Recommended Dilutions:
WB IP IP IF-Cell |
1:5,000-1:25,000 1:5,000-1:10,000 (C-FLAG) 1:1,000-1:5,000 (N-FLAG) 1:250 |
| Alternative names: | DDDDK epitope tag DDDK ddk DYKDDDDK DYKDDDDK epitope tag DYKDDDDK tag ECS epitope tag ECS tag Enterokinase Cleavage Site epitope tag Enterokinase Cleavage Site tag FLAG FLAG tag |
Images
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Fig1:
Western blot analysis of DYKDDDDK Tag ( FLAG ) on different lysates with Mouse anti-DYKDDDDK Tag ( FLAG ) antibody (HA601080) at different dilutions. Lane 1/2/3: C-terminal FLAG-tag fusion protein lysate Lane 4/5/6: N-terminal FLAG-tag fusion protein lysate Lysates/proteins at 50 ng/Lane. Observed band size: 35/36 kDa Exposure time: 2 minutes; 12% 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 (HA601080) at different dilutions was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:150,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of HeLa cells labeling DYKDDDDK Tag ( FLAG ) with Mouse anti-DYKDDDDK Tag ( FLAG ) antibody (HA601080) at 1/250 dilution. HeLa cells, transfected with FLAG-tagged empty control, Claudin 18.2 (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-DYKDDDDK Tag ( FLAG ) antibody (HA601080) at 1/250 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. beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution. |
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Fig3:
FLAG-tag was immunoprecipitated in 2µg C-terminal FLAG-tag fusion protein lysate with HA601080. Western blot was performed from the immunoprecipitate using HA601080 at 1/5,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1:2,000 dilution was used for 60 mins at room temperature. Lane 1: C-terminal FLAG-tag fusion protein lysate (input). Lane 2: HA601080 IP in C-terminal FLAG-tag fusion protein lysate. Lane 3: Mouse IgG instead of HA601080 IP in C-terminal FLAG-tag fusion protein lysate. Blocking/Dilution buffer: 5% NFDM/TBST. |
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Fig4:
FLAG-tag was immunoprecipitated in 2µg N-terminal FLAG-tag fusion protein lysate with HA601080. Western blot was performed from the immunoprecipitate using HA601080 at 1/5,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1:2,000 dilution was used for 60 mins at room temperature. Lane 1: N-terminal FLAG-tag fusion protein lysate (input). Lane 2: HA601080 IP in N-terminal FLAG-tag fusion protein lysate. Lane 3: Mouse IgG instead of HA601080 IP in N-terminal FLAG-tag fusion protein lysate. Blocking/Dilution buffer: 5% NFDM/TBST. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.