ChIP answers Flashcards
what does ChIP stand for
Chromatin Immunoprecipitation
what year was ChIP developed
1984
who developed ChIP
John T. Lis, and David Gilmour
what are the two types of ChIP
Native (N-ChIP), and Crosslinked (X-ChIP)
Native ChIP
involves the digestion of native chromatin by a nuclease without the use of a fixing agent
Crosslink ChIP
involves chemical fixation but precipitation is less efficient
what is ChIP used for (3)
1) to investigate protein-DNA interactions
2) to look at the state of chromatin and gene transcription
3) to understand gene expression and regulation in cells and tissues of interest
5 main steps of ChIP
1) cross-linking and lysis,
2) chromatin fragmentation
3) immunoprecipitation
4) DNA purification
5) DNA quantification and analysis
cross linkig and lysis
proteins in the chromatin are crosslinked, and then cells are lysed using SDS containing buffer
chromatin fragmentation
chromatin is cut into smaller fragments to isolate DNA-protein complexes and antibodies are attached
immuneprecipitation
antibodies for the protein of interest are attached to the DNA-Protein complexes and form precipitation at the bottom of the solution
purification
crosslinking is reversed and purification occurs to isolate complexes from the DNA
DNA quantification and analysis
results are quantified using ChIP-qPCR or ChIP sequencing
