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Immunohistochemical analysis of paraffin-embedded human brain tissue using E2F4 recombinant monoclonal antibody, clone 24E12 (Cat # RAB07574) 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 Goat anti-rabbit IgG polymer labeled by HRP and visualized using 0.05% DAB.

Immunofluorescent staining of Hela Cells with E2F4 recombinant monoclonal antibody, clone 24E12 (Cat # RAB07574), counter-stained with DAPI. The cells were fixed in 4% formaldehyde 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 513-congugated AffiniPure Goat Anti-Rabbit IgG(H+L).

Flow cytometry shows MCF7 cells stained with E2F4 recombinant monoclonal antibody, clone 24E12 (Cat # RAB07574)(red line). The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/1*10⁶cells) for 45min at 4°C. The secondary antibody used was FITC-conjugated Goat Anti-rabbit IgG(H+L) at 1:200 dilution for 35min at 4°C. Control antibody (green line) was rabbit IgG (1ug/1*10⁶cells) used under the same conditions. Acquisition of >10,000 events was performed.