Western Blot
Positive WB detected in: V5-tagged fusion protein 1, 2, 3, 4, 5, 6 at 50ng
All lanes: V5 antibody at 1:2000
Secondary
Goat polyclonal to mouse IgG at 1/50000 dilution
Predicted band size: 31.9, 20.4, 55, 24.2, 53.4, 30 KDa
Observed band size: 32, 21, 55, 28, 58, 33 KDa
Exposure time: 5min

Western Blot
Positive WB detected in: 50ng V5-tagged fusion protein
V5 Tag antibody at 1:10000, 1:20000, 1:40000, 1:80000, 1:160000, 1:320000, 1:640000, 1:1280000
Secondary
Goat polyclonal to mouse IgG at 1/50000 dilution
Predicted band size: 32 KDa
Observed band size: 32 KDa
Exposure time: 5min

Western Blot
Positive WB detected in: Mouse anti V5 tag Monoclonal antibody at 1:2000
Lane 1: Recombinant V5-tagged fusion protein at 25ng
Lane 2: Recombinant V5-tagged fusion protein at 12.5ng
Lane 3: Recombinant V5-tagged fusion protein at 6.25ng
Lane 4: Recombinant V5-tagged fusion protein at 3.125ng
Lane 5: Recombinant V5-tagged fusion protein at 1.56ng
Secondary
Goat polyclonal to Mouse IgG at 1/50000 dilution
Predicted band size: 32 kd
Observed band size: 32 kd
Exposure time: 5min

Immunofluorescence staining of transfected HEK293F cells with CSB-MA000161M1m at 1:100, counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 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 R-PE Goat Anti-Mouse IgG(H+L).

Immunoprecipitating V5 Tag in Transfected HEK293F cells whole cell lysate
Lane 1: Mouse control IgG2a instead of CSB-MA000161M1m in Transfected HEK293 cells whole cell lysate
Lane 2: CSB-MA000161M1m (2.5μl) + Transfected HEK293F cells whole cell lysate (500μg)
Lane 3: Transfected HEK293F cells whole cell lysate (20μg)
For western blotting, the blot was detected with CSB-MA000161M1m at 1:3000, and a HRP-conjugated Protein G antibody was used as the secondary antibody at 1:50000