Chromatin Immunoprecipitation (ChIP) Protocol using Protein A beads (Dorsky Lab)
Shan-Fu's Fu’s adaptation from Kazuyuki's Kazuyuki’s protocol (Grunwald Lab) (8/24/05)
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(Comments: This protocol is not for ChIP-Seq since other types of carrier DNA (such as calf thymus DNA) were used for blocking. Washing and blocking protein A beads are necessary for eliminating non-specific beads' beads’ binding.)
<Day 1>
¨ Cross-linking of protein and DNA
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(Comments: The amount of embryos needed for ChIP need to be optimized by specific sample. It also depends on your antibody specificity. As far as I know, I can see difference between negative control and sample with only 10 embryos using dynabeads. But I haven't haven’t check it with protein A beads.
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Remove sup. (Beads from no-antibody tube could serve as "no Ab" “no Ab” negative control after washing and elution)
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