Active CCAAT/Enhancer Binding Protein Gamma (CEBPg)

CCAAT/Enhancer Binding Protein Gamma (CEBPg), a member of the C/EBP family of basic leucine zipper transcription factors, is encoded by the CEBPG gene in humans and widely expressed in various tissues and cell types. It regulates gene transcription by binding to CCAAT/enhancer elements in promoter regions, participating in multiple cellular processes including cell proliferation, differentiation, metabolism and immune response. CEBPg also forms homo- or heterodimers with other C/EBP family members, and its dysregulation is associated with the pathogenesis of several diseases such as cancer and inflammatory disorders. Notably, CEBPg physically interacts with ATF4, a stress-responsive transcription factor, to form functional heterodimers that modulate the expression of target genes under cellular stress.To detect the activity of recombinant human CEBPg, a functional ELISA assay was performed to evaluate the interaction between recombinant human CEBPg and recombinant human ATF4. Briefly, biotin-linked CEBPg were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100µl were then transferred to ATF4-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µl stop solution to the wells and read at 450nm immediately. The binding activity of human CEBPg and human ATF4 was shown in Figure 1, the EC50 for this effect is 0.202µg/mL.