| Application | Recommended Dilution |
|---|---|
| WB | 1:1000-1:5000 |
| ICC | 1:10-1:100 |
| IF | 1:10-1:100 |
| IP | 1:200-1:2000 |
Western Blot
Detected samples: Hela whole cell lysate, 293 whole cell lysate, A549 whole cell lysate, K562 whole cell lysate; Untreated (-) or treated (+) with 30mM sodium butyrate for 4h
All lanes: HIST1H3A antibody at 1:2000
Secondary
Goat polyclonal to rabbit IgG at 1/40000 dilution
Predicted band size: 16 kDa
Observed band size: 16 kDa
Immunocytochemistry analysis of CSB-PA010418OA23butHU diluted at 1:20 and staining in Hela cells (treated with 30mM sodium butyrate 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.
Immunofluorescence staining of Hela cells (treated with 30mM sodium butyrate for 4h) with CSB-PA010418OA23butHU at 1:10, 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 HIST1H3A in Hela whole cell lysate
Lane 1: Rabbit control IgG instead of CSB-PA010418OA23butHU 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-PA010418OA23butHU (5µg) + Hela whole cell lysate (500µg)
Lane 3: Hela whole cell lysate (20µg)
Chromatin Immunoprecipitation Hela (106, treated with 30mM sodium butyrate for 4h) were treated with Micrococcal Nuclease, sonicated, and immunoprecipitated with 5µg anti-HIST1H3A (CSB-PA010418OA23butHU) 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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