VALIDATION IMAGES
MCF-7 lysates probed with AKT1 (11F2) Monoclonal Antibody (bsm-52010R) at 1:1000 overnight at 4˚C. Followed by a conjugated secondary antibody.
Lane 1: Mouse Cerebrum lysates; Lane 2: Mouse Thymus lysates; Lane 3: Mouse Heart lysates; Lane 4: Mouse Lung lysates; Lane 5: Mouse NIH/3T3 cell lysates; Lane 6: Rat Cerebellum lysates; Lane 7: Rat Thymus lysates; Lane 8: Rat Heart lysates; Lane 9: Human MCF-7 cell lysates; Lane 10: Human Hela cell lysates; Lane 11: Human U937 cell lysates; Lane 12: Human A549 cell lysates probed with AKT1 (11F2) Monoclonal Antibody, Unconjugated (bsm-52010R) 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 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 AKT1 (11F2) Monoclonal Antibody, Unconjugated (bsm-52010R) 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 AKT1 (11F2) Monoclonal Antibody, Unconjugated (bsm-52010R) at 1:200 overnight at 4°C, 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 AKT1 (11F2) Monoclonal Antibody, Unconjugated (bsm-52010R) at 1:200 overnight at 4°C, DAB staining.
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 AKT1 (11F2) Monoclonal Antibody, Unconjugated (bsm-52010R) 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 AKT1 (11F2) Monoclonal Antibody, Unconjugated (bsm-52010R) 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 AKT1 (11F2) Monoclonal Antibody, Unconjugated (bsm-52010R) at 1:200 overnight at 4°C, 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 AKT1 (11F2) Monoclonal Antibody, Unconjugated (bsm-52010R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AKT1 Monoclonal Antibody, Unconjugated(bsm-52010R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AKT1 Monoclonal Antibody, Unconjugated(bsm-52010R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AKT1 Monoclonal Antibody, Unconjugated(bsm-52010R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AKT1 Monoclonal Antibody, Unconjugated(bsm-52010R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
25 ug total protein per lane of various lysates (see on figure) probed with AKT1 monoclonal antibody, unconjugated (bsm-52010R) at 1:5000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.