Crotonyl-HIST1H3A (K9) Antibody

Western Blot
Detected samples: Hela whole cell lysate, Jurkat whole cell lysate, HepG2 whole cell lysate; Untreated (-) or treated (+) with 30mM sodium crotonylate for 4h
All lanes: HIST1H3A antibody at 1:100
Secondary
Goat polyclonal to rabbit IgG at 1/50000 dilution
Predicted band size: 16 kDa
Observed band size: 16 kDa

Immunocytochemistry analysis of CSB-PA010418OA09crHU diluted at 1:100 and staining in Hela cells (treated with 30mM sodium crotonylate for 4h) performed on a Leica BondTM system. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

Immunoprecipitating HIST1H3A in Hela whole cell lysate (treated with 30mM sodium crotonylate for 4h)
Lane 1: Rabbit control IgG instead of CSB-PA010418OA09crHU in Hela whole cell lysate (treated with 30mM sodium crotonylate for 4h). For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
Lane 2: CSB-PA010418OA09crHU (5µg) + Hela whole cell lysate (treated with 30mM sodium crotonylate for 4h) (500µg)
Lane 3: Hela whole cell lysate (treated with 30mM sodium crotonylate for 4h) (20µg)

Chromatin Immunoprecipitation Hela (10⁶, treated with 30mM sodium crotonylate for 4h) were treated with Micrococcal Nuclease, sonicated, and immunoprecipitated with 5µg anti-HIST1H3A (CSB-PA010418OA09crHU) or a control normal rabbit IgG. The resulting ChIP DNA was quantified using real-time PCR with primers against the β-Globin promoter.