Flag Tag Monoclonal Antibody

Western Blot
Positive WB detected in: 10ng Flag Tag fusion protein Flag Tag antibody at 1:100000, 1:200000, 1:400000, 1:800000, 1:1600000
Secondary
Goat polyclonal to mouse IgG at 1/50000 dilution
Predicted band size: 35 kDa
Observed band size: 35 kDa
 

Western Blot
Positive WB detected in: Flag Tag fusion protein at 50ng, 25ng, 12.5ng, 6.25ng, 3.625ng, 1.5625ng, 0.78ng
All lanes: Flag Tag antibody at 1:1000
Secondary
Goat polyclonal to mouse IgG at 1/50000 dilution
Predicted band size: 35 kDa
Observed band size: 35 kDa
 

Western Blot
Positive WB detected in: Flag Tag fusion protein 1, 2, 3, 4 at 50ng
All lanes: Flag Tag antibody at 1:1000
Secondary
Goat polyclonal to mouse IgG at 1/50000 dilution
Predicted band size: 59.6, 36, 35, 42 kDa
Observed band size: 59.6, 36, 35, 42 kDa
 

Western Blot
Positive WB detected in: Flag Tag fusion protein at 10ng
All lanes: Company A, Company B, Company C, Company D, CSB-MA000021M0m antibody at 1:50000, 1:100000, 1:200000, 1:400000, 1:800000, 1:1600000, 1:3200000, 1:6400000
Secondary
Goat polyclonal to Mouse IgG at 1/50000 dilution
Predicted band size: 35 kDa
Observed band size: 35 kDa
 

Western Blot
Positive WB detected in: Flag Tag fusion protein at 50ng, 25ng, 12.5ng, 6.25ng, 3.125ng, 1.5625ng, 0.78ng, 0.39ng
All lanes: Company A, Company B, Company C, Company D, CSB-MA000021M0m antibody at 1:1000
Secondary
Goat polyclonal to Mouse IgG at 1/50000 dilution
Predicted band size: 35 kDa
Observed band size: 35 kDa
 

Western Blot
Positive WB detected in: 5 different recombinant proteins with Flag Tag
All lanes: Company A, Company B, Company C, Company D, CSB-MA000021M0m antibody at 1:5000
Secondary
Goat polyclonal to Mouse IgG at 1/50000 dilution
 

Immunoprecipitating Flag Tag in Transfected HEK-293 cell whole cell lysate
Lane 1: Mouse control IgG instead of CSB-MA000021M0m in Transfected HEK-293 cell whole cell lysate
Lane 2: CSB-MA000021M0m (1µl) + Transfected HEK-293 cell whole cell lysate (500µg)
Lane 3: Transfected HEK-293 cell whole cell lysate (20µg)
For western blotting, the blot was detected with CSB-MA000021M0m at 1:5000, and a HRP-conjugated Protein G antibody was used as the secondary antibody at 1:2000
 

Immunoprecipitating Flag Tag in 293F transfected whole cell lysate
Lane 1: Mouse control IgG instead of CSB-MA000021M0m in Flag Tag in 293F transfected whole cell lysate. For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
Lane 2: Company A (1µl), Company B (1µl), Company C (1µl), Company D (1µl), CSB-MA000021M0m (1µl) + Flag Tag in 293F transfected whole cell lysate (500µg)
Lane 3: Flag Tag in 293F transfected whole cell lysate (20µg)

Immunofluorescence staining of 293T cells transfected with Flag tag by CSB-MA000021M0m at 1:300, counter-stained with DAPI. The cells were fixed in 4% formaldehyde and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L). The image on the right is the 293T cells transfected without Flag tag.