H4K20me1 Polyclonal Antibody

To check the specificity of H4K20me1 Polyclonal Antibody (bs-53035R) at 1:20,000 dilution, a Dot Blot was performed with peptides containing different modifications of histone H4 and the unmodified sequence. 0.2 to 100 pmol concentration of peptides were spotted on to a membrane. Experiment shows specificity of the antibody for the modification of interest.

To determine the titer, an ELISA was performed using a serial dilution of the H4K20me1 Polyclonal Antibody (bs-53035R), to detect peptide containing histone modification of interest. By plotting the absorbance against antibody dilutions, titer of the antibody was estimated to be 1:8,000.

ChIP were performed using U2OS cells using H4K20me1 Polyclonal Antibody (bs-53035R). Chromatin was sheared from 1.6 million cells. A titration of the antibody consisting of 2, 5, 10 and 15\u03bcl per ChIP experiment was analyzed. IgG (5\u03bcg\/IP) was used as negative IP control. Quantitative PCR was performed with primers for the GAPDH promoter and for the coding region of the myogenic differentiation gene (MYOD) which is inactive at normal conditions. This figure shows the recovery, expressed as a % of input.