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Chromatin Immunoprecipitation (ChIP) assay is an important method for transcriptional regulation monitoring with uncovered knowledge of interactions between specific proteins and a genomic DNA region. Given the fact that DNA-binding proteins (including transcription factors and histones) in living cells can be cross-linked to the DNA that they are binding, ChIP is used to immunoprecipitate the protein–DNA complex out of cellular lysates by an appropriate antibody. n

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chromatin immunoprecipitation protocol

Chromatin Immunoprecipitation Protocol

Chromatin Immunoprecipitation (ChIP) assay is an

transcriptional regulation monitoring with uncovered knowledge of interactions

between specific proteins and a genomic DNA region. Given the fact that

DNA-binding proteins (including transcription factors and histones) in living cells can

be cross-linked to the DNA that they are binding, ChIP is used to immunoprecipitate

the protein–DNA complex out of cellular lysates by an appropriate antibody. The

immunoprecipitated complexes are collected by centrifugation, and proteins are

eluted for further analysis such as western blot and LC/MS. Nucleic acid analysis is

achieved through PCR, RT-PCR, cDNA sequencing and microarray. This protocol

provides a detailed procedure to achieve successful ChIP assay in a fast and efficient

manner.

important method for

Reagents:

37% formaldehyde

1 M glycine

Cell lysis buffer: 150 mM NaCl, 50 mM Tris-HCl (pH 7.5), 5 mM EDTA, 0.5% NP-40, 1%

Triton X-100, with 1× protease inhibitor cocktail.

PBS: pH 7.4, 137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 2 mM KH2PO4.

10% Chelex 100

https://www.creative-diagnostics.com/chromatin-immunoprecipitation-chip-protoco

l.htm