H4K5ac Polyclonal Antibody

Dot Blot analysis was performed with peptides containing a variety of histone modifications and the unmodified H4. 0.2-100 pmol of the respective peptides were spotted on a membrane. H4K5ac Polyclonal Antibody (bs-53099R) was used at 1:5,000 dilution. Result shows high specificity of the antibody for the modification of interest.

ChIP were performed using HeLa cells using H4K5ac Polyclonal Antibody (bs-53099R). Chromatin was sheared from 1 million cells. A titration of the antibody consisting of 1, 2, 5, and 10\u03bcg per ChIP experiment was analyzed. IgG (2\u03bcg\/IP) was used as negative IP control. Quantitative PCR was performed with primers for promoter of the active gene EIF4A2 and for a region 1 kb upstream of the GAPDH as positive controls, and for the inactive MYOD1 gene and Sat2 satellite repeat region used as negative controls. Result shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

ELISA was performed using a serial dilution of the\r\nH4K5ac Polyclonal Antibody (bs-53099R) against peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:8,900.

HeLa cells were stained with the H4K5ac Polyclonal Antibody (bs-53099R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K5ac antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two staining is shown on the right.