RT Journal Article
SR Electronic
T1 Systematic comparison of monoclonal versus polyclonal antibodies for mapping histone modifications by ChIP-seq
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 054387
DO 10.1101/054387
A1 Michele Busby
A1 Cathy Xue
A1 Yossi Farjoun
A1 Elizabeth Gienger
A1 Ido Yofe
A1 Adrianne Gladden
A1 Chad Nusbaum
A1 Alon Goren
YR 2016
UL http://biorxiv.org/content/early/2016/05/19/054387.abstract
AB The robustness of ChIP-seq datasets is highly dependent upon the antibodies used. Currently, polyclonal antibodies are the standard despite several limitations: they are non-renewable, vary in performance between lots, and need to be validated with each new lot. In contrast, monoclonal antibody lots are renewable and provide consistent performance. To increase ChIP-seq standardization, we investigated whether monoclonal antibodies could replace polyclonal antibodies. We compared monoclonal antibodies that target five key histone modifications (H3K4me1, H3K4me3, H3K9me3, H3K27ac and H3K27me3) to their polyclonal counterparts. Overall performance was highly similar for four monoclonal/polyclonal pairs. In contrast, the binding patterns for H3K27ac differed substantially between polyclonal and monoclonal antibodies. However, this was most likely due to the distinct immunogen used rather than the clonality of the antibody. Altogether, we found that monoclonal antibodies as a class perform as well as polyclonal antibodies. Accordingly, we recommend the use of monoclonal antibodies in ChIP-seq experiments.