VALIDATION IMAGES
Formalin-fixed and paraffin embedded rat brain labeled with Anti-Phospho-PERK(Thr980) Polyclonal Antibody, Unconjugated (bs-3330R) at 1:200 followed by conjugation to the secondary antibody and DAB staining
Formalin-fixed and paraffin-embedded human lung carcinoma labeled with Anti-Phospho-PERK(Thr980) Polyclonal Antibody, Unconjugated(bs-3330R) 1:200, overnight at 4°C, The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(bs-0295G-Cy3)used at 1:200 dilution for 40 minutes at 37°C.
Paraformaldehyde-fixed, paraffin embedded rat kidney tissue; Antigen retrieval by boiling in sodium citrate buffer(pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (normal goat serum) at 37°C for 20min; Antibody incubation with Rabbit Anti-PERK (Thr980) Polyclonal Antibody, Unconjugated (bs-3330R) at 1:400 overnight at 4°C, followed by a conjugated secondary and DAB staining
Paraformaldehyde-fixed, paraffin embedded Rat pancreas; 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 PERK(Thr980) Polyclonal Antibody, Unconjugated (bs-3330R) at 1:200 overnight at 4°C, DAB staining.
A431 cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with
PERK(Thr980) Polyclonal Antibody(bs-3330R)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(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
K562 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 Phospho-PERK (Thr980) Polyclonal Antibody(bs-3330R)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 (human pancreas); 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 (PERK (Thr980)) Polyclonal Antibody, Unconjugated (bs-3330R) 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 pancreas); 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 (PERK (Thr980)) Polyclonal Antibody, Unconjugated (bs-3330R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
HepG2 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 Phospho-PERK (Thr980) Antibody(bs-3330R)at 1:100 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).
Lane 1: Human MCF-7 cell lysates; Lane 2: Human MCF-7-Tg cell lysates; Lane 3: Human MCF-7-phosphatase cell lysates probed with Phospho-PERK (Thr980) Polyclonal Antibody, Unconjugated (bs-3330R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.