ChIP was performed on macrophages derived from mouse bone marrow using the ab against PPARG and optimized PCR primer sets for qPCR. Sheared chromatin from 1 million cells and 1 ug of PPARg antibody were used per ChIP experiment. IgG was used as a negative IP control. Figure 1A: recovery, expressed as the % of input, of the PDK4 PPAR response element (RE). Figure 1B: recovery of the FABP4 Adipo PPAR RE in cells treated with RSG, a very strong activating ligand of PPARG, and in untreated cells.
Determination of the titer To determine the titer, an ELISA was performed using a serial dilution of the antibody against human PPARG. The plates were coated with the peptide used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:70, 250.