Chromatin Immunoprecipitation Validation Protocol

ChIP assays were performed with H1299 cells in accordance with
standard protocols^{43 (link)}. Normal rabbit IgG served as the control. ChIP samples were
analyzed by quantitative real-time PCR using the FastStart Universal SYBR Green
Master (Roche). A standard curve was prepared for each set of primers using serial
titration of the input DNA. The percentage of ChIP DNA was calculated relative to
the input DNA from primer-specific standard curves using the Rotor-Gene 6000
Series Software 1.7. The primer sequences for ChIP are listed in Supplementary
Table 4. Antibodies used were: H4S1ph
(Abcam; ab14723), H4K5ac (Millipore; CS204381), H4R3me2a (Active Motif; 39705),
H4R3me2s (Abcam; ab5823), CK2α (Abcam; ab70774), H3K4me3 (Abcam; ab8580), and
H3K27me3 (Abcam; ab6002).

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Ju J., Chen A., Deng Y., Liu M., Wang Y., Wang Y., Nie M., Wang C., Ding H., Yao B., Gui T., Li X., Xu Z., Ma C., Song Y., Kvansakul M., Zen K., Zhang C.Y., Luo C., Fang M., Huang D.C., Allis C.D., Tan R., Zeng C.K., Wei J, & Zhao Q. (2017). NatD promotes lung cancer progression by preventing histone H4 serine phosphorylation to activate Slug expression. Nature Communications, 8, 928.

Publication 2017

FastStart Universal SYBR GreenMaster H4K5ac H4R3me2a ab5823 ab70774 ab8580 ab6002

Antibodies Cells Chip Chip assays Ck2α Dna primer Gene H3k4me3 Primer Quantitative real time pcr Rabbit

Corresponding Organization :

Other organizations : Nanjing University, Southeast University, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Walter and Eliza Hall Institute of Medical Research, University of Melbourne, La Trobe University, Rockefeller University, Nanjing University of Chinese Medicine

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Protocol cited in 2 other protocols

Variable analysis

independent variables

H1299 cells

dependent variables

Percentage of ChIP DNA relative to input DNA

control variables

Standard protocols for ChIP assays
Normal rabbit IgG as control

controls

Normal rabbit IgG

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