| Application | Recommended Dilution |
|---|---|
| WB | 1:500-1:2000 |
| ICC | 1:20-1:200 |
| IP | 1:200-1:2000 |
Western Blot
Detected samples: Hela whole cell lysate, Jurkat whole cell lysate, 293 whole cell lysate; Untreated (-) or treated (+) with 10mM sodium propionate 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
Immunoprecipitating HIST1H4A in Hela whole cell lysate
Lane 1: Rabbit control IgG instead of CSB-PA010429OA05propHU in Hela whole cell lysate. For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
Lane 2: CSB-PA010429OA05propHU (5µg) + Hela whole cell lysate (500µg)
Lane 3: Hela whole cell lysate (20µg)
Immunocytochemistry analysis of CSB-PA010429OA05propHU diluted at 1:100 and staining in Hela cells (treated with 10mM sodium propionate 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.
Chromatin Immunoprecipitation Hela (4*106, treated with 10mM sodium propionate for 4h) were treated with Micrococcal Nuclease, sonicated, and immunoprecipitated with 5µg anti-HIST1H4A (CSB-PA010429OA05propHU) or a control normal rabbit IgG. The resulting ChIP DNA was quantified using real-time PCR with primers against the β-Globin promoter.
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