Histone H3 [ac Lys9] Antibody (PSH04-47)

Immunohistochemistry: Histone H3 [ac Lys9] Antibody (PSH04-47) [NBP3-32430]

Immunohistochemistry: Histone H3 [ac Lys9] Antibody (PSH04-47) [NBP3-32430] - Immunohistochemical analysis of paraffin-embedded human stomach tissue with Rabbit anti-Histone H3 antibody (NBP3-32430) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunocytochemistry/ Immunofluorescence: Histone H3 [ac Lys9] Antibody (PSH04-47) [NBP3-32430]

Immunocytochemistry/ Immunofluorescence: Histone H3 [ac Lys9] Antibody (PSH04-47) [NBP3-32430] - Immunocytochemistry analysis of C6 cells labeling Histone H3 with Rabbit anti-Histone H3 antibody (NBP3-32430) at 1/10,000 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Histone H3 antibody at 1/10,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.

Flow Cytometry: Histone H3 [ac Lys9] Antibody (PSH04-47) [NBP3-32430]

Flow Cytometry: Histone H3 [ac Lys9] Antibody (PSH04-47) [NBP3-32430] - Flow cytometric analysis of HeLa cells labeling Histone H3.

Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32430, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Western Blot: Histone H3 [ac Lys9] Antibody (PSH04-47) [NBP3-32430]

Western blot analysis on different lysates with NBP3-32430 at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 500ng/mL TSA for 4 hours cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: NIH/3T3 treated with 400nM TSA for 18 hours cell lysate Lane 5: C6 cell lysate Lane 6: C6 treated with 1μM TSA for 18 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 15 kDa Observed band size: 15 kDa Exposure time: 6 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. NBP3-32430 at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG [HRP] Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.