Blank control: A431. Primary Antibody (green line): Rabbit Anti-Histone H2B antibody (TC52099) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-AF488 Dilution: 1μg /test. Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

Blank control: Hela. Primary Antibody (green line): Rabbit Anti-Histone H2B antibody (TC52099) Dilution: 1:50; Secondary Antibody : Goat anti-rabbit IgG-AF488 Dilution: 1:1000. Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (human tonsil); 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 (Histone H2B) Monoclonal Antibody, Unconjugated (TC52099) at 1:50 overnight at 4°C, followed by operating according to SignalEnhance IHC KIT（Rabbit）Cat#A10026 instructions and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human breast 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 (Histone H2B) Monoclonal Antibody, Unconjugated (TC52099) at 1:50 overnight at 4°C, followed by operating according to SignalEnhance IHC KIT（Rabbit）Cat#A10026 instructions and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse 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 (Histone H2B) Monoclonal Antibody, Unconjugated (TC52099) at 1:50 overnight at 4°C, followed by operating according to SignalEnhance IHC KIT（Rabbit）Cat#A10026 instructions and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse liver); 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 (Histone H2B) Monoclonal Antibody, Unconjugated (TC52099) at 1:50 overnight at 4°C, followed by operating according to SignalEnhance IHC KIT（Rabbit）Cat#A10026 instructions and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse testis); 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 (Histone H2B) Monoclonal Antibody, Unconjugated (TC52099) at 1:50 overnight at 4°C, followed by operating according to SignalEnhance IHC KIT（Rabbit）Cat#A10026 instructions and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human liver); 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 (Histone H2B) Monoclonal Antibody, Unconjugated (TC52099) at 1:50 overnight at 4°C, followed by operating according to SignalEnhance IHC KIT（Rabbit）Cat#A10026 instructions and DAB staining.

A431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (Histone H2B) monoclonal Antibody, Unconjugated (TC52099) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Sample: Lane 1: Hela cell lysates Lane 2: PC-12 cell lysates Primary: Anti-Histone H2B (TC52099) at 1/500 dilution Secondary: Goat Anti-Rabbit IgG - HRP at 1/5000 dilution Predicted band size: 14 kD Observed band size: 14 kD