VALIDATION IMAGES
hepg2 lysates probed with AMPK alpha-1 (Thr198) Polyclonal Antibody, Unconjugated (bs-5551M) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:10000 for 60 min at 37˚C.
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 AMPK alpha-1 (Thr198) Polyclonal Antibody, Unconjugated (bs-5551R) at 1:400 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat heart; 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 AMPK alpha-1 (Thr198) Polyclonal Antibody, Unconjugated (bs-5551R) 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 AMPK alpha-1 (Thr198) Polyclonal Antibody, Unconjugated (bs-5551R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human pancreatic 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 AMPK alpha-1 (Thr198) Polyclonal Antibody, Unconjugated (bs-5551R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat heart; 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 AMPK alpha-1 (Thr198) Polyclonal Antibody, Unconjugated (bs-5551R) at 1:200 overnight at 4°C, DAB staining.
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 AMPK alpha-1 (Thr198) Polyclonal Antibody, Unconjugated (bs-5551R) 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 AMPK alpha-1 (Thr198) Polyclonal Antibody, Unconjugated (bs-5551R) at 1:200 overnight at 4°C, DAB staining.
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 AMPK alpha-1 (Thr198) Polyclonal Antibody, Unconjugated (bs-5551R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse heart; 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 AMPK alpha-1 (Thr198) Polyclonal Antibody, Unconjugated (bs-5551R) at 1:200 overnight at 4°C, DAB staining.
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 AMPK alpha-1 (Thr198) Polyclonal Antibody, Unconjugated (bs-5551R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse liver; 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 AMPK alpha-1 (Thr198) Polyclonal Antibody, Unconjugated (bs-5551R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat liver; 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 AMPK alpha-1 (Thr198) Polyclonal Antibody, Unconjugated (bs-5551R) at 1:200 overnight at 4°C, DAB staining.
U937 cells 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 AMPK alpha-1 (Thr198) Antibody(bs-5551R)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).
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 (phospho-AMPK alpha-1 (Thr183)) Polyclonal Antibody, Unconjugated (bs-5551R) 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 colon carcinoma); 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 (phospho-AMPK alpha-1 (Thr183)) Polyclonal Antibody, Unconjugated (bs-5551R) 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 kidney); 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 (phospho-AMPK alpha-1 (Thr183)) Polyclonal Antibody, Unconjugated (bs-5551R) 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-AMPK alpha-1 (Thr183) Antibody(bs-5551R)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).