Histone H3 (tri methyl K27) Recombinant Rabbit Monoclonal Antibody [PSH28-13]

☑ Cell treatment (CT)

Western blot analysis of Histone H3 (tri methyl K27) on different lysates with Rabbit anti-Histone H3 (tri methyl K27) antibody (HA724754) at 1/2,000 dilution.

Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
Lane 2: SH-SY5Y (Human neuroblastoma cell) cell lysate
Lane 3: HCT 116 (Human colon cancer cell) cell lysate
Lane 4: NIH/3T3 (Mouse fibroblast) cell lysate
Lane 5: C6 (Rat glioma cell) cell lysate
Lane 6: MCF7 (Human breast cancer cell) cell lysate
Lane 7: MCF7 treated with 20μM EED226 for 72 hours cell lysate

Lysates/proteins at 20 µg/Lane.
Exposure time: 21 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724754, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 15 kDa
Observed band size: 15 kDa

Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA724754, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

Application: Immunocytochemistry (IF-cell)

Species: Mouse
Sample: NIH/3T3 (Mouse fibroblast)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA724754, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

Application: Immunocytochemistry (IF-cell)

Species: Rat
Sample: C6 (Rat glioma cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA724754, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).