QuikChIP (with Loading Control) Kit (ChIP)

Chromatin Immunoprecipitation: QuickChip (with Loading Control) ChIP Kit [NBP2-29497] - Lane 1: 100 bp ladder. Lane 2: 293 cells - RNApol II - GAPDH primers. Lane 3: Hela cells - RNApol II - GAPDH primers. Lane 4: A549 cells - RNApol II - GAPDH primers. Lane 5: 293 cells - RNApol II - Neg control primers. Lane 6: HeLa cells - RNApol II - Neg control primers. Lane 7: A549 cells - RNApol II - Neg control primers.

• Applications: ChIP

QuikChIP (with Loading Control) Kit (ChIP) Summary

• Description: 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 enough DNA for five ChIP assays.
  
  Features:
  
• Quick and simple.
• Optimized ready-to-use buffers and protocol are included.
• Can be used for both histone and non-histone proteins.
• Kit Type: Kit (ChIP)

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP)

Packaging, Storage & Formulations

• Storage: Storage of components varies. See protocol for specific instructions.

Alternate Names for QuikChIP (with Loading Control) Kit (ChIP)

• Quik ChIP
• QuikChIP

Background

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 nonhistone proteins from the chromatin binding site, it is necessary to incorporate a cross-linking step. This 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 non-histone 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 mmunoprecipitated 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.

Limitations

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 (with Loading Control) Kit (NBP2-29497)(3)

Publications using NBP2-29497

• Vinothkumar K, Chanda S, Singh VK et al. EVI1 upregulates PTGS1 (COX1) and decreases the action of tyrosine kinase inhibitors (TKIs) in chronic myeloid leukemia cells International journal of hematology 2022-10-25 [PMID: 36282419] (Chemotaxis)
• Cao Y, Feng Z, He X Et al. Prolactin-regulated Pbk is involved in pregnancy-induced beta cell proliferation in mice The Journal of endocrinology 2021-11-01 [PMID: 34788229]
• Massoumi R, Kuphal S, Hellerbrand C et al. Down-regulation of CYLD expression by Snail promotes tumor progression in malignant melanoma. J Exp Med. 2009-01-16 [PMID: 19124656] (Chemotaxis)
  
  Details:
  30101K (Quick ChIP Kit), ChIP assay demonstrated Snail1 to the CYLD promoter. Table 3S, Fig 2A: primary melanoma cells, and melanoma cells.