Mouse anti human c ebpβ antibody

Immunoprecipitation was conducted to reveal the protein interaction between C/EBPβ and E2F2 [23 (link)]. About 2 × 10⁶ cells were collected, and protein A/G agarose beads were used, which captured IgG from virtually all isotypes in cell culture supernatants and other antibodies for efficient antibody purification to remove nonspecific proteins in the first step. Then, the supernatants were incubated overnight at 4 °C with mouse anti-human C/EBPβ antibody (1:1000, santa cruz); afterward, protein A/G agarose beads were used again to catch C/EBPβ binding proteins and their protein conjugates. Subsequently, the aforementioned protein complexes were eluted from the beads, and then, agarose gel electrophoresis and Western blot were performed to transfer target proteins to PVDF membranes, followed by incubating with mouse anti-human C/EBPβ antibody (1:1000, santa cruz) or mouse anti-human E2F2 antibody (1:500, santa cruz). Next, the specific secondary antibody (Goat Anti-Mouse IgG HRP, 1:10,000, abmart), eliminating heavy chain interference, was used. Finally, the immunoblots were detected using an ECL detection system and quantified by Image J software (Version No.: 3.2.0.8).