DYKDDDDK tag Rabbit Monoclonal Antibody(ARA797)

Predicted MW: Depend on fusion protein with DYKDDDDK tag
Lane 1: 293 cells lysate transfected with C-terminal DYKDDDDK tagged gene (ARA797 at 1:20,000 dilution).
Lane 2: 293 cells lysate transfected with N-terminal DYKDDDDK tagged gene (ARA797 at 1:10,000 dilution).
Lane 3: 293 cells lysate without any transfection (ARA797 at 1:2,000 dilution).
Lane 4: Multi-tag fusion protein (ARA797 at 1:2,000 dilution)
Lane 1/2/3: 3 µg per lane
Lane 4: 20 ng per lane
2nd Ab:
GAR HRP(H+L) 1:5,000

DYKDDDDK tag was immunoprecipitated from 0.1mg of 293 whole cells lysate transfected with N-terminal DYKDDDDK tagged gene with ARA797 at 1:50 dilution.
2nd Ab:
GAR HRP for IP 1:500
Lane 1: ARA797 IP in 293 whole cell lysate transfected with N-terminal DYKDDDDK tagged gene
Lane 2: PBS instead of ARA797 in 293 whole cell lysate transfected with N-terminal DYKDDDDK tagged gene
Lane 3: 293 whole cell lysate transfected with N-terminal DYKDDDDK tagged gene, 2 µg (input)
Exposure: 30s

ARA797 staining DYKDDDDK tag in 293 cells transfected with N-terminal DYKDDDDK tagged gene by IF/ICC (immunofluorescence/immunocytochemistry). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 10% goat serum for half an hour at room temperature. Samples were incubated with primary antibody (1:10,000) at 4°C. An Alexa Fluor^® 488-conjugated Goat Anti-Rabbit IgG polyclonal was used as the secondary antibody (1:500). DAPI (blue) was used as the nuclear counter stain.

Control: PBS and secondary antibody, An Alexa Fluor^® 488-conjugated Goat Anti-Rabbit IgG (1:500).

ARA797 staining DYKDDDDK tag in 293 cells transfected with C-terminal DYKDDDDK tagged gene by IF/ICC (immunofluorescence/immunocytochemistry). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 10% goat serum for half an hour at room temperature. Samples were incubated with primary antibody (1:10,000) at 4°C. An Alexa Fluor^® 488-conjugated Goat Anti-Rabbit IgG polyclonal was used as the secondary antibody (1:500). DAPI (blue) was used as the nuclear counter stain.

Control: PBS and secondary antibody, An Alexa Fluor^® 488-conjugated Goat Anti-Rabbit IgG (1:500).

Overlay histogram showing 293 cells transfected with N-terminal DYKDDDDK tagged gene stained with ARA797 (Red). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% TritonX-100 for 15 min. The cells were then incubated in the antibody (ARA797, 1:2,000 dilution) in 1x PBS/1% BSA for 30 min at room temperature. The secondary antibody used was a Goat Anti-Rabbit Alexa Fluor^® 488 (IgG H+L) at 1:2,000 dilution for 20 min at room temperature. Unlabelled sample (Black) was used as a control.

Overlay histogram showing 293 cells transfected with C-terminal DYKDDDDK tagged gene stained with ARA797 (Red). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% TritonX-100 for 15 min. The cells were then incubated in the antibody (ARA797, 1:2,000 dilution) in 1x PBS/1% BSA for 30 min at room temperature. The secondary antibody used was a Goat Anti-Rabbit Alexa Fluor^® 488 (IgG H+L) at 1:2,000 dilution for 20 min at room temperature. Unlabelled sample (Black) was used as a control.