We do not recommend using antibodies that fail Luminex and/or SNAP-ChIP criteria for ChIP-seq experiments, as the data will be contaminated by a significant amount of off-target signal, which could alter biological analyses and conclusions. “Limited Data” describes antibodies that show specificity, but were only tested against a small subset of nucleosomes. These antibodies should also be avoided for ChIP-seq.
* Denotes this antibody was tested in Shah et al. 2018.
SNAP-ChIP qPCR Data: The antibody was tested in a native ChIP experiment using chromatin from K-562 cells, with EpiCypher SNAP-ChIP Spike-ins added prior to micrococcal nuclease digestion. Specificity (Y-axis) was determined by qPCR for the DNA barcodes corresponding to designer nucleosomes in the SNAP-ChIP Panel (X-axis). Data is normalized to relevant on-target nucleosome. For K-AcylStat panel: tetraAc-H3 is K4,K9,K14,K18ac; tetraAc-H4 is K5,K8,K12,K16ac; tetraAc-H2A is K5,K8,K13,K15ac.
Luminex Data: In this assay, biotinylated modified recombinant nucleosomes from EpiCypher (X-axis) were individually coupled to uniquely identifiable Luminex® MagPlex® beads. Antibody binding to nucleosomes was tested in multiplex and detected using a species-specific anti-IgG PE on the Luminex FlexMAP 3D®. Data is normalized to relevant on-target nucleosome.