Mouse Anti-V5-Tag Recombinant Antibody (clone 8A7A8) (CAT#: ZG-0275U)

Figure 1 Mouse Anti-V5-Tag Antibody (ZG-0275U) in WB

Positive WB detected in: V5-tagged fusion protein 1, 2, 3, 4 at 50ng All lanes: V5 antibody at 1: 2000
Secondary
Goat polyclonal to mouse IgG at 1/50000 dilution
Predicted band size: 50.2, 47.6, 66.7, 37.8 KDa
Observed band size: 52, 47, 67, 42 KDa
Exposure time: 30s

Figure 2 Mouse Anti-V5-Tag Antibody (ZG-0275U) in IF

Immunofluorescence staining of transfected HEK293F cells with ZG-0275U 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).

Figure 3 Mouse Anti-V5-Tag Antibody (ZG-0275U) in FC

Overlay histogram showing untransfected HEK293F cells (green line) or transfected HEK293F cells (red line) stained with ZG-0275U. The cells were fixed with 70% Ethylalcohol (18h) and then permeabilized with 0.3% Triton X-100 for 2 min. The cells were then incubated in 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1:200/1*106cells) for 1 h at 4°C. The secondary antibody used was FITC goat anti-mouse IgG(H+L) at 1/100 dilution for 30min at 4°C. Acquisition of>10,000 events was performed.

Figure 4 Mouse Anti-V5-Tag Antibody (ZG-0275U) in IP

Immunoprecipitating V5 Tag in Transfected HEK293F cells whole cell lysate Lane 1: Mouse control IgG1 in Transfected HEK293F cells whole cell lysate
Lane 2: ZG-0275U (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 ZG-0275U at 1:5000, and a HRP-conjugated Protein G antibody was used as the secondary antibody at 1:50000