Lactyl-Histone H3 (K18) Recombinant Rabbit Monoclonal Antibody [PSH26-77]

☑ Cell treatment (CT)

Western blot analysis of Lactyl-Histone H3 (K18) on different lysates with Rabbit anti-Lactyl-Histone H3 (K18) antibody (HA724642) at 1/5,000 dilution.

Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
Lane 2: HeLa treated with 100mM Lactate sodium for 24 hours cell lysate
Lane 3: HT-29 (Human colorectal adenocarcinoma cell) cell lysate
Lane 4: HT-29 treated with 100mM Lactate sodium for 24 hours cell lysate

Lysates/proteins at 20 µg/Lane.
Exposure time: 3 minutes; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724642, 1/5,000 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

☑ Cell treatment (CT)

Western blot analysis of Lactyl-Histone H3 (K18) on different lysates with Rabbit anti-Lactyl-Histone H3 (K18) antibody (HA724642) at 1/5,000 dilution.

Lane 1: NIH/3T3 (Mouse fibroblast) cell lysate
Lane 2: NIH/3T3 treated with 100mM Lactate sodium for 24 hours cell lysate

Lysates/proteins at 15 µg/Lane.
Exposure time: 42 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724642, 1/5,000 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

☑ Cell treatment (CT)

Immunoprecipitation (IP)

Lactyl-Histone H3 (K18) was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) treated with 100mM Lactate sodium for 24 hours cell lysate with HA724642 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724642 at 1/5,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa treated with 100mM Lactate sodium for 24 hours cell lysate (input)
Lane 2: HA724642 IP in HeLa treated with 100mM Lactate sodium for 24 hours cell lysate
Lane 3: Rabbit IgG instead of HA724642 in HeLa treated with 100mM Lactate sodium for 24 hours cell lysate

Exposure time: 3 minutes
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA724642, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

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

Dot blot analysis of Lactyl-Histone H3 (K18) on different peptides with Rabbit anti-Lactyl-Histone H3 (K18) antibody (HA724642) at 1/5,000 dilution.

Lane 1: Lactyl-Histone H3 (Lys18)
Lane 2: Acetyl-Histone H3 (Lys18)
Lane 3: Unmodified Histone H3

Proteins loading: 100ng, 50ng, 10ng, 5ng;
Exposure time: 20 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724642, 1/5,000 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