Rabbit Anti-ATF4 Recombinant Antibody (clone 9E1) (CAT#: ZG-0553J)

Figure 1 Rabbit Anti-ATF4 Antibody (ZG-0553J) in WB

Western Blot
Positive WB detected in: Hela whole cell lysate, MCF-7 whole cell lysate, 293 whole cell lysate, HepG2 whole cell lysate, Jurkat whole cell lysate, K562 whole cell lysate
All lanes: ATF4 antibody at 1.6μg/ml
Secondary
Goat polyclonal to rabbit IgG at 1/50000 dilution
Predicted band size: 39 KDa
Observed band size: 50 KDa

Figure 2 Rabbit Anti-ATF4 Antibody (ZG-0553J) in IHC

IHC image of ZG-0553J diluted at 1:160 and staining in paraffin-embedded human breast cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

Figure 3 Rabbit Anti-ATF4 Antibody (ZG-0553J) in IF

Immunofluorescence staining of HepG2 cells with ZG-0553J at 1:53, 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 488-congugated Goat Anti-Rabbit IgG (H+L).

Figure 4 Rabbit Anti-ATF4 Antibody (ZG-0553J) in IP

Immunoprecipitating ATF4 in Hela whole cell lysate
Lane 1: Rabbit control IgG instead of ZG-0553J in Hela whole cell lysate. For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
Lane 2: ZG-0553J (3μg) + Hela whole cell lysate (500μg)
Lane 3: Hela whole cell lysate (20μg)

Figure 5 Rabbit Anti-ATF4 Antibody (ZG-0553J) in FC

Overlay histogram showing Hela cells stained with ZG-0553J (red line) at 1:50. 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 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by primary antibody for 1 h at 4°C. The secondary antibody used was FITC goat anti-rabbit IgG (H+L) at 1/200 dilution for 1 h at 4°C. Control antibody (green line) was used under the same conditions. Acquisition of>10,000 events was performed.