Butyrly-HIST1H4A (K5) Antibody

Western Blot
Detected samples: Hela whole cell lysate, Jurkat whole cell lysate, 293 whole cell lysate, HepG2 whole cell lysate; Untreated (-) or treated (+) with 30mM sodium butyrate for 4h
All lanes: HIST1H4A antibody at 1:1000
Secondary
Goat polyclonal to rabbit IgG at 1/40000 dilution
Predicted band size: 12 kDa
Observed band size: 12 kDa

Immunofluorescence staining of HepG2 cells (treated with 30mM sodium butyrate for 4h) with CSB-PA010429OA05butHU, counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L).

Immunoprecipitating HIST1H4A in HepG2 whole cell lysate (treated with 30mM sodium butyrate for 4h)
Lane 1: Rabbit control IgG instead of CSB-PA010429OA05butHU in HepG2 whole cell lysate (treated with 30mM sodium butyrate for 4h). For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
Lane 2: CSB-PA010429OA05butHU (5μg) + HepG2 whole cell lysate (treated with 30mM sodium butyrate for 4h) 500μg)
Lane 3: HepG2 whole cell lysate (treated with 30mM sodium butyrate for 4h) (20μg)

Chromatin Immunoprecipitation Hela (4*10⁶, treated with 30mM sodium butyrate for 4h) were treated with Micrococcal Nuclease, sonicated, and immunoprecipitated with 5μg anti-HIST1H4A (CSB-PA010429OA05butHU) or a control normal rabbit IgG. The resulting ChIP DNA was quantified using real-time PCR with primers against the β-Globin promoter.