VALIDATION IMAGES
Lane 1:Mouse embryo lysate Lane 2:Mouse eye lysate probed with Rabbit Anti-ATF4/CREB-2 Polyclonal Antibody, Unconjugated (bs-1531R) at 1:300 overnight at 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:5000 for 90 min at RT.
Formalin-fixed and paraffin embedded mouse brain labeled with Anti-ATF4/CREB-2 Polyclonal Antibody, Unconjugated (bs-1531R) at 1:200 followed by conjugation to the secondary antibody and DAB staining
Formalin-fixed and paraffin embedded human rectal carcinoma labeled with Anti-ATF4/CREB-2 Polyclonal Antibody, Unconjugated (bs-1531R) at 1:200 followed by conjugation to the secondary antibody and DAB staining
Paraformaldehyde-fixed, paraffin embedded Rat spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with ATF4/CREB-2 Polyclonal Antibody, Unconjugated (bs-1531R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with ATF4/CREB-2 Polyclonal Antibody, Unconjugated (bs-1531R) at 1:200 overnight at 4°C, DAB staining.
Lane 1: Rat Cerebrum lysates; Lane 2: Mouse Cerebrum lysates probed with ATF4/CREB-2 Polyclonal Antibody, Unconjugated (bs-1531R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Paraformaldehyde-fixed, paraffin embedded Rat lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with ATF4/CREB-2 Polyclonal Antibody, Unconjugated (bs-1531R) at 1:200 overnight at 4°C, DAB staining.
Mouse spleen cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with ATF4 Antibody(bs-1531R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with ATF4/CREB-2 Polyclonal Antibody, Unconjugated (bs-1531R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with ATF4/CREB-2 Polyclonal Antibody, Unconjugated (bs-1531R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human breast cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with ATF4/CREB-2 Polyclonal Antibody, Unconjugated (bs-1531R) at 1:200 overnight at 4°C, DAB staining.
Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (ATF4) polyclonal Antibody, Unconjugated (bs-1531R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Lane 1: Mouse Spleen Lysates; Lane 2: Mouse Lung Lysates; Lane 3: Mouse Brain Lysates; Lane 4: Rat Brain Lysates; Lane 5: Human Hela cell Lysates; Lane 6: Human HepG2 cell Lysates. Probed with ATF4 polyclonal Antibody, unconjugated (bs-1531R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
U2OS cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (ATF4) polyclonal Antibody, Unconjugated (bs-1531R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Paraformaldehyde-fixed, paraffin embedded (rat skin); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (ATF4/CREB-2) Polyclonal Antibody, Unconjugated (bs-1531R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse skin); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (ATF4/CREB-2) Polyclonal Antibody, Unconjugated (bs-1531R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Human skin cancer); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (ATF4/CREB-2) Polyclonal Antibody, Unconjugated (bs-1531R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Human breast cancer); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (ATF4/CREB-2) Polyclonal Antibody, Unconjugated (bs-1531R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Human brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (ATF4/CREB-2) Polyclonal Antibody, Unconjugated (bs-1531R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Lane 1: Mouse Cerebrum tissue lysates; Lane 2: Rat Cerebrum tissue lysates; Lane 3: Human HeLa cell lysates; Lane 4: Human Jurkat cell lysates; Lane 5: Human HL-60 cell lysates probed with ATF4 Polyclonal Antibody, Unconjugated (bs-1531R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.