PPARg polyclonal antibody (sample size)

ChIP results obtained with the Diagenode antibody directed against PPARg
ChIP was performed with the Diagenode antibody against PPARg (Cat. No. C15410367) on sheared chromatin from 4,000,000 Capan2 cells with the iDeal ChIP-seq kit for TF’s. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the HIST1H4C and LAMP1 genes, used as positive controls, and for the MYOD1 gene, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

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ChIP-seq results obtained with the Diagenode antibody directed against PPARg
ChIP was performed on sheared chromatin from 4,000,000 Capan2 cells using 5 µg of the Diagenode antibody against PPARg (Cat. No. C15410367) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 1.5 Mb region of chromosome 1 (figure 2A and B), in a genomic region of chromosome 6 containing several HIST1 genes (figure 2C), and in a genomic region of the X-chromosome surrounding the LAMP1 positive control gene (figure 2D).

Determination of the antibody titer
To determine the titer of the antibody, an ELISA was performed using a serial dilution of Diagenode antibody directed against PPARg (Cat. No. C15410367). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be >1:1,000,000.