Paraformaldehyde-fixed, paraffin embedded mouse brain; 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 AMPK alpha-1/2 (Thr183/Thr172) Antibody (bs-4002R) at 1:400 overnight at 4°C, followed by a conjugated secondary and DAB staining.


HepG2 cell lysates probed with phospho-AMPK alpha-2 (Thr172) Polyclonal Antibody, Unconjugated (bs-4002R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.


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 AMPK alpha-1/2 (Thr183/Thr172) Antibody, ALEXA FLUOR® 647 Conjugated , FITC Conjugated Antibody（bs-4002R-FITC） at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green) and isotype control (orange).


A549 cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 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-1/2 (Thr183/Thr172) Antibody(bs-4002R-AF488)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).


Mouse cells(black) 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 AMPK alpha-1/2 (Thr183/Thr172) Antibody(bs-4002R-AF488)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).


Lane 1: Mouse Cerebral cortex lysates probed with AMPK alpha-1/2 (Thr183/Thr172) Polyclonal Antibody, Unconjugated (bs-4002R) 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 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-1/2 (Thr183/Thr172) Polyclonal Antibody, Unconjugated (bs-4002R) 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-1/2 (Thr183/Thr172) Polyclonal Antibody, Unconjugated (bs-4002R) 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-1/2 (Thr183/Thr172) Polyclonal Antibody, Unconjugated (bs-4002R) 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-1/2 (Thr183/Thr172) Polyclonal Antibody, Unconjugated (bs-4002R) at 1:200 overnight at 4°C, DAB staining.


Lane 1: Mouse Eye lysates; Lane 2: Rat Muscle lysates; Lane 3: Rat Heart lysates; Lane 4: Rat Kidney lysates; Lane 5: Rat Eye lysates probed with AMPK alpha-1/2 Polyclonal Antibody, Unconjugated (bs-4002R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C


U-937 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-1/2 (Thr183/Thr172) Antibody(bs-4002R)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).