Anti-Bak1 antibody

Fig1: Tissue/cell: human colon carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;; Incubation: Anti-Bak Polyclonal Antibody, Unconjugated 1:200, overnight at 4℃, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Fig2: Sample:Lung (Mouse) Lysate at 30 ug; Primary: Anti-Bak at 1:300 dilution;; Secondary: HRP conjugated Goat-Anti-Rabbit IgG(bse-0295G) at 1: 5000 dilution;; Predicted band size : 23kD; Observed band size : 23kD

Fig3: Tissue/cell: human lung carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;; Incubation: Anti-Bak Polyclonal Antibody, Unconjugated 1:500, overnight at 4℃, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Fig4: Blank control: Hela(blue).; Primary Antibody:Rabbit Anti- Bak antibody(bs-1638R), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;; Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions );; Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.; Protocol; The cells were fixed with 2% paraformaldehyde (10 min) , then permeabilized with 90% ice-cold methanol for 30 min on ice. Antibody ( 1μg /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody of 175212# at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.