DYKDDDDK Tag ( FLAG ) Mouse Monoclonal Antibody [A2-A4]
cat.: M1403-2
| Product Type: | Mouse monoclonal IgG1, primary antibodies |
|---|---|
| Species reactivity: | Species independent |
| Applications: | WB, IP, ELISA, IF-Cell |
| Clonality: | Monoclonal |
| Clone number: | A2-A4 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 2ug/ul |
| Purification: | Protein G affinity purified. |
| Isotype: | IgG1 |
| Immunogen: | Synthetic peptide immune sequence is N-DYKDDDDK-C. |
| Positive control: | DYKDDDDK Tag recombinant protein, C-terminal FLAG-tagged recombinant protein, N-terminal FLAG-tagged recombinant protein. |
| Recommended Dilutions:
WB IP IF-Cell |
1:5,000-1:10,000 2-5 µg/ml. 1:100 |
| 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
|
Fig1:
Western blot analysis of DYKDDDDK Tag on Flag-tagged recombinant protein Flag-Ki67 and Ki67-Flag at different cell lysates level. Predicted band size: 35 kDa; Observed band size: 35 kDa; Exposure time: 60 seconds; SDS-PAGE gel concentration:12% Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (M1403-2, 1/2,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:200,000 dilution was used for 1 hour at room temperature. |
|
Fig2:
Western blot analysis of DYKDDDDK Tag on Flag-tagged recombinant protein 2xflag-Histone H3.1 at different dilutions of HeLa cells with anti-DYDDDDK antibody (M1403-2). Lane 1: Non-transfected HeLa cell lysates (1/2000) Lane 2~4: Transfected HeLa cell lysates (1/2500; 1/5000; 1/10,000) Predicted band size: 15 kDa; Observed band size: 18 kDa; Exposure time: 30 seconds; SDS-PAGE gel concentration:12%. Cell lysate at 10ug per lane. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (M1403-2) was used at different dilutions in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:200,000 dilution was used for 1 hour at room temperature. |
|
Fig3:
Immunoprecipitation analysis of N-Flag in HeLa cells. Lane 1: HeLa transfected with DYKDDDDK-tagged human Histone H3.1 expression vector whole cell lysate (input). Lane 2: M1403-2 IP in HeLa transfected with DDDDK-tagged human Histone H3.1 expression vector whole cell lysate. Lane 3: Mouse IgG (HA1027) instead of M1403-2 in HeLa transfected with DYKDDDDK-tagged human Histone H3.1 expression vector whole cell lysate. Predicted band size: 19 kDa; Observed band size: 19 kDa; Exposure time: 60 seconds; SDS-PAGE gel concerntration: 15%. DYKDDDDK Tag was immunoprecipitated from 0.5 mg Hela transfected with human 2x Flag- Histone H3.1 expression vector whole cell lysate with M1403-2 2 ug/mL. Western blot was performed from the immunoprecipitate using M1403-2 at 1/5,000 dilution for 1 hour at room temperature. The secondary antibody for IP detection reagent, Goat anti-Mouse IgG-HRP antibody (HA1006), was used at 1:100,000 dilution. |
|
Fig4:
Immunoprecipitation analysis of C-Flag in HeLa cells. Lane 1: HeLa transfected with DYKDDDDK-tagged human Siglec15 expression vector whole cell lysate (input). Lane 2: M1403-2 IP in HeLa transfected with DDDDK-tagged human Siglec15 expression vector whole cell lysate. Lane 3: Mouse IgG (HA1027) instead of M1403-2 in HeLa transfected with DYKDDDDK-tagged human Siglec15 expression vector whole cell lysate. DYKDDDDK Tag was immunoprecipitated from 0.5 mg Hela transfected with human Siglec15 expression vector whole cell lysate with M1403-2 2 ug/mL. Western blot was performed from the immunoprecipitate using M1403-2 at 1/2,000 dilution for 1 hour at room temperature. The secondary antibody for IP detection reagent, Goat anti-Mouse IgG-HRP antibody (HA1006), was used at 1:100,000 dilution. |
|
Fig5:
Immunofluorescent analysis of 2xFlag tagged Histone H3.1 in HeLa cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling DYKDDDDK tag with M1403-2 at 1/100 dilution, followed by iFluor™ 488 Goat anti-mouse IgG antibody (HA1125) at 1/1000 dilution (green). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.