Chromatin Immunoprecipitation Hela (4*106) were treated with Micrococcal Nuclease, sonicated, and immunoprecipitated with 5µg anti-H2AFZ (CSB-PA010100OA04nacHU) or a control normal rabbit IgG. The resulting ChIP DNA was quantified using real-time PCR with primers against the beta-Globin promoter.
Immunofluorescence staining of Hela cells with CSB-PA010100OA04nacHU 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).
IHC image of CSB-PA010100OA04nacHU diluted at 1:50 and staining in paraffin-embedded human prostate tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was 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.
IHC image of CSB-PA010100OA04nacHU diluted at 1:50 and staining in paraffin-embedded human brain tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was 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.
Western Blot Positive WB detected in: K562 whole cell lysate, Mouse spleen tissue All lanes: H2AFZ antibody at 1µg/ml Secondary Goat polyclonal to rabbit IgG at 1/50000 dilution Predicted band size: 14 kDa Observed band size: 14 kDa
Immunoprecipitating H2AFZ in K562 whole cell lysate Lane 1: Rabbit control IgG (1µg) instead of CSB-PA010100OA04nacHU in K562 whole cell lysate. For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000) Lane 2: CSB-PA010100OA04nacHU (8µg) + K562 whole cell lysate (500µg) Lane 3: K562 whole cell lysate (10µg)
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