Chromatin Immunoprecipitation: Quick ChIP Kit [NBP2-29902] - Human MCF-7 cells were fixed for 10 minutes with 1% formaldehyde. The cells were lysed and the chromatin sonicated (200-1000 bp fragments). Chromatin was ...read more
The QuikChIP(TM) Kit contains components for 25 ChIP assays optimized for use with transcriptionally active chromatin in adherent or suspension mammalian cells. The protocol is optimized for use with one 10 cm plate containing 5 x 10^6 cells per preparation. Each ChIP assay requires approximately 1 x 10^6 cells. Thus, a 10 cm plate (one preparation) of cells will provide DNA sufficient for five ChIP assays.
Quick and simple.
Optimized ready-to-use buffers and protocol are included.
Can be used for both histone and non-histone proteins.
Reagents for 25 ChIP assays and sheared DNA preparations.
Use in Immunoprecipitation reported in scientific literature (PMID 21957289).
Read Publications using NBP2-29902 in the following applications:
Mouse reactivity reported in scientific literature (PMID: 23709694). Bacteria reactivity reported in scientific literature (PMID: 24363348). Human reactivity reported in scientific literature (PMID: 21086175).
Packaging, Storage & Formulations
Storage of components varies. See protocol for specific instructions.
0.5M EDTA (250 uL)
100X PIC (2 x 0.75 mL)
100X PMSF (2 x 0.75 mL)
10X Glycine (25 mL)
10X PBS (55 mL)
1M NaHCO3 (1.25 mL)
1M Tris-HCl pH 6.5 (500 uL)
20% SDS (625 uL)
5M NaCl (500 uL)
ChIP Dilution Buffer (50 mL)
Proteinase K at 10 mg/mL (60 uL)
RNase A at 10 mg/mL (60 uL)
Salmon Sperm DNA/Protein A/G Agarose (2 x 1.75 mL)
SDS Lysis Buffer (25 mL)
Wash Buffer A (25 mL)
Wash Buffer B (25 mL)
Wash Buffer C (25 mL)
Wash Buffer D (25 mL)
Additional Materials Required (not included in this kit): Reagents: Cells of interest, Antibody of interest, Appropriate antibody control, 37% Formaldehyde, 3M Sodium Acetate (pH 5.2), Phenol/chloroform (optional), 100% ethanol, 70% ethanol, Taq DNA polymerase, dNTP mixture, Cell culture media, ddH2O. Equipment: Agarose gel electrophoresis apparatus, Rotating platform, Microcentrifuge, Microcentrifuge tubes, Spectrophotometer, Variable volume pipettors, Pipette tips, Sonicator, Thermocycler, Thermocycling tubes, Cell Scraper, Aspirator.
Chromatin Immunoprecipitation (ChIP) is a powerful method used to identify regions of the genome associated with specific proteins. These associations are crucial for vital cellular functions including gene transcription, DNA replication and recombination, repair, segregation, chromosomal stability, cell cycle progression, and epigenetic silencing. The ChIP assay has been used to study both histones and non-histone proteins, such as transcription factors, within the context of the cell. Transcription factors and other DNA binding proteins have a weaker affinity than histones, which generally are tightly associated within the chromatin complex. To avoid dissociation of non-histone proteins from the chromatin binding site, it is necessary to incorporate a cross-link step. QuikChIP(TM) Kit has been optimized for use with both histone and non-histone proteins. Using the QuikChIP(TM) Assay, cells are fixed with formaldehyde to cross-link nonhistone proteins to DNA. Alternatively, the cross-linking steps are not needed for histone proteins, which are tightly bound to DNA. The chromatin is released from the nuclei and sheared into 200-1000 bp fragments. The chromatin is then immunoprecipitated with specific antibodies. DNA sequences cross-linked to the protein of interest co-precipitate as part of the chromatin complex. The DNA/Chromatin/Antibody complex is then isolated using Protein G Agarose. After reverse cross-linking, the associated DNA is released from the complex and is ready for analysis.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.
Publications for QuikChIP Kit (NBP2-29902)(46)
We have publications tested in 6 confirmed species: Human, Mouse, Rat, Bacteria, Bovine, Insect.
We have publications tested in 2 applications: ChIP, IP.