VALIDATION IMAGES
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 PTEN Polyclonal Antibody, Unconjugated (bs-0686R) at 1:500 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 PTEN Polyclonal Antibody, Unconjugated (bs-0686R) at 1:800 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse thyroid; 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 PTEN Polyclonal Antibody, Unconjugated (bs-0686R) at 1:200 overnight at 4°C, DAB staining.
Hela 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 PTEN Antibody(bs-0686R)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 Kidney tissue lysates; Lane 2: Rat Kidney tissue lysates probed with PTEN Polyclonal Antibody, Unconjugated (bs-0686R) 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 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; Incubation with (PTEN) Polyclonal Antibody, Unconjugated (bs-0686R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
25 ug total protein per lane of various lysates (see on figure) probed with PTEN polyclonal antibody, unconjugated (bs-0686R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.