VALIDATION IMAGES
Human A549 lysates probed with Pan-Actin Polyclonal Antibody, Unconjugated (bs-10196R) at 1:300 overnight at 4˚C. Followed by a conjugated secondary antibody (bs-0295G-HRP) at 1:5000 for 90 min at RT.
HeLa cells probed with Pan-Actin Polyclonal Antibody, unconjugated (bs-10196R) at 1:100 dilution for 30 minutes compared to control cells (dark blue), secondary only (light blue) and isotype control (orange)
Lane 1: Rat Liver lysates; Lane 2: Rat Urinary bladder lysates; Lane 3: Mouse Stomach lysates; Lane 4: A549 cell lysates probed with Pan-Actin Polyclonal Antibody, Unconjugated (bs-10196R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: A549 cell lysates; Lane 2: NIH/3T3 cell lysates; Lane 3: Hela cell lysates; Lane 4: A431 cell lysates ; Lane 5: HepG2 cell lysates;Lane 6: Mouse Stomach lysates; Lane 7: Mouse Lung lysates; Lane 8: Mouse Skeletal muscle (-) lysates probed with Pan-Actin Polyclonal Antibody, Unconjugated (bs-10196R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
NIH/3T3 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 Pan-Actin Polyclonal Antibody(bs-10196R)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: Mouse Lung lysates; Lane 2: Human HepG2 cell lysates; Lane 3: Mouse Small intestine lysates probed with Pan-Actin Polyclonal Antibody, Unconjugated (bs-10196R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Human A549 cell lysates; Lane 2: Mouse NIH/3T3 cell lysates; Lane 3: Human Hela cell lysates; Lane 4: Human A431 cell lysates; Lane 5: Human HepG2 cell lysates; Lane 6: Mouse Stomach lysates; Lane 7: Mouse Lung lysates; Lane 8: Mouse Skeletal muscle lysates probed with Pan-Actin Polyclonal Antibody, Unconjugated (bs-10196R) 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 (Human stomach); 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 (Pan-Actin) Polyclonal Antibody, Unconjugated (bs-10196R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat stomach); 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 (Pan-Actin) Polyclonal Antibody, Unconjugated (bs-10196R) 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 stomach); 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 (Pan-Actin) Polyclonal Antibody, Unconjugated (bs-10196R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat colon); 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 (Pan-Actin) Polyclonal Antibody, Unconjugated (bs-10196R) 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 (Pan-Actin) Polyclonal Antibody, Unconjugated (bs-10196R) 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 cervical 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 (Pan-Actin) Polyclonal Antibody, Unconjugated (bs-10196R) 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 duodenum); 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 (Pan-Actin) Polyclonal Antibody, Unconjugated (bs-10196R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.