Immunocytochemistry analysis of CSB-PA010389OA13acHU diluted at 1:5 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-PA010389OA13acHU at 1:2.5, 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).
Western Blot Positive WB detected in: Hela whole cell lysate, 293 whole cell lysate, A549 whole cell lysate, Jurkat whole cell lysate, HepG2 whole cell lysate (all treated with 30mM sodium butyrate for 4h) All lanes: HIST1H2AG antibody at 1.25µg/ml Secondary Goat polyclonal to rabbit IgG at 1/50000 dilution Predicted band size: 15 kDa Observed band size: 15 kDa
Immunoprecipitating HIST1H2AG in 293 whole cell lysate Lane 1: Rabbit control IgG instead of CSB-PA010389OA13acHU in 293 whole cell lysate. For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000) Lane 2: CSB-PA010389OA13acHU (5µg) + 293 whole cell lysate (500µg) Lane 3: 293 whole cell lysate (20µg)
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