Chromatin Accessibility Assay Kit (ab185901)
Key features and details
- Assay type: Quantitative
- Assay time: 1 hr 30 min
- Sample type: Adherent cells, Suspension cells, Tissue
製品の概要
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製品名
Chromatin Accessibility Assay Kit -
サンプルの種類
Tissue, Adherent cells, Suspension cells -
アッセイタイプ
Quantitative -
全工程の試験時間
1h 30m -
種交差性
交差種: Mammals -
製品の概要
Chromatin Accessibility Assay Kit ab185901 is a complete set of optimized reagents designed for conducting a gene-specific analysis of chromatin accessibility including nucleosome/transcription factor positioning from various biological samples via real time PCR.
The chromatin accessibility assay works on the principle that accessible chromatin is easily accessed by a nuclease mix to digest the DNA. When the DNA from the nuclease-treated sample and a no-nuclease control is analyzed for a particular gene by PCR, the more accessible the chromatin at that gene, the lower the signal of the nuclease-treated sample relative to the no-nuclease control.
Chromatin accessibility assay protocol summary:
- lyze cells and extract chromatin by adding lysis buffer to cell pellet, resuspending and incubating for 10 min
- centrifuge at 5,000 rpm for 5 min, discard supernatant, resuspend chromatin pellet in wash buffer and centrifuge again
- resuspend chromatin pellet in nuclease reaction mix (or no-nuclease control mix to controls) and incubate for 4 min
- add stop solution and incubate for 10 min
- add Proteinase K and incubate for 60 min
- add samples to DNA binding columns, spin, and discard flow through
- add DNA washing solution to columns, spin, and discard flow through 3 times
- add elution solution and spin to elute DNA
- analyze gene target with PCR and compare nuclease-treated to no-nuclease control -
特記事項
The accessibility of regulatory elements in chromatin is critical for many aspects of gene regulation. Nucleosomes positioned over regulatory elements inhibit access of transcription factors to DNA. To elucidate the role of the interactions between chromatin and transcription factors, it is crucial to determine chromatin accessibility through mapping of the nucleosome positioning along the genome. In general, the more condensed the chromatin, the more difficult it is for transcription factors and other DNA binding proteins to access DNA and carry out their tasks. The more accessible the DNA, the more likely surrounding genes are actively transcribed. The presence (or the absence) of nucleosomes directly or indirectly affects a variety of other cellular and metabolic processes such as recombination, replication, centromere formation, and DNA repair.
製品の特性
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保存方法
Please refer to protocols. -
内容 48 tests 1000X Protease Inhibitor Cocktail 1 x 50µl 10X DNA Washing Solution 1 x 3ml 10X Lysis Buffer 1 x 11ml 10X Wash Buffer 1 x 10ml DNA Binding Solution 1 x 15ml Elution Solution 1 x 1ml F-Collection Tube 1 x 50 units F-Spin Column 1 x 50 units Negative Control Primer-F (20 μM) 1 x 10µl Negative Control Primer-R (20 μM) 1 x 10µl Nuclear Digestion Buffer 1 x 3ml Nuclease Mix 1 x 50µl Positive Control Primer-F (20 μM) 1 x 10µl Positive Control Primer-R (20 μM) 1 x 10µl Proteinase K (10 mg/mL) 1 x 110µl Reaction Stop Solution 1 x 0.5ml -
研究分野
画像
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Chromatin accessibility of opened chromatin (euchromatin)
Amplification of proximal promoter regions for the constitutively expressed target gene was carried out in MDA-231 cells by using positive control primers. Red and green lines: Nse-untreated; Blue lines: Nse-treated. The difference of Ct values between Nse-treated and untreated is > 7 cycles.
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Chromatin accessibility of closed chromatin (heterochromatin)Amplification of proximal promoter regions for the constitutively repressed target gene was carried out in MDA-231 cells by using negative control primers. Red and green lines: Nse-untreated; Blue and pink lines: Nse-treated. The difference of Ct values between Nse-treated and untreated is <1.5 cycles.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (7)
ab185901 は 7 報の論文で使用されています。
- Hari-Gupta Y et al. Myosin VI regulates the spatial organisation of mammalian transcription initiation. Nat Commun 13:1346 (2022). PubMed: 35292632
- Li T et al. Dynamic nucleosome landscape elicits a noncanonical GATA2 pioneer model. Nat Commun 13:3145 (2022). PubMed: 35672415
- Osburn SC et al. Effects of end-stage osteoarthritis on markers of skeletal muscle Long INterspersed Element-1 activity. BMC Res Notes 15:245 (2022). PubMed: 35799274
- Lindqvist B et al. Chromatin maturation of the HIV-1 provirus in primary resting CD4+ T cells. PLoS Pathog 16:e1008264 (2020). PubMed: 31999790
- Romero MA et al. Five months of voluntary wheel running downregulates skeletal muscle LINE-1 gene expression in rats. Am J Physiol Cell Physiol 317:C1313-C1323 (2019). PubMed: 31618076
- Mumford PW et al. Skeletal muscle LINE-1 retrotransposon activity is upregulated in older versus younger rats. Am J Physiol Regul Integr Comp Physiol 317:R397-R406 (2019). PubMed: 31188650
- Pentz R et al. Cardioprotective cytokine interleukin-33 is up-regulated by statins in human cardiac tissue. J Cell Mol Med 22:6122-6133 (2018). PubMed: 30216659