Pierce Agarose ChIP Kit / 琼脂糖ChIP试剂盒
货号：26156
规格：30 reactions
价格：9072
产品类型：免疫印迹/组化
品牌：Thermo Fisher
| The Thermo Scientific Pierce Agarose ChIP Kit provides a complete set of reagents and a simple, fast and reproducible protocol to perform chromatin immunoprecipitation (ChIP) assays.The Pierce Agarose ChIP Kit contains sufficient reagents to perform 30 ChIP assays with appropriate controls using an optimized protocol and a convenient spin-column format. ChIP-validated and quality-guaranteed antibodies are also available for use the the Pierce Agarose ChIP Kit. Because antibodies that work for western blotting may not function in ChIP assays, we have assembled our most popular ChIP-validated monoclonal and polyclonal antibodies here to help you move from experimental setup to results more quickly.The strength of the ChIP assay is its ability to capture a snapshot of specific protein-DNA interactions as they occur in living cells, and then quantitate the interactions using standard or quantitive PCR. A successful ChIP assays requires a number of critical steps to be carried out (crosslinking, chromatin preparation, immunoprecipitation) prior to detecting the target genomic DNA sequence. Individually, each step in the ChIP assay protocol can be time consuming to optimize. The Pierce Agarose ChIP Assay Kit simplifies the process, making accurate and reproducible results easier to obtain.To perform ChIP assays with the Pierce Agarose ChIP Kit, protein-DNA complexes are stabilized and then extracted. In vivo crosslinking is traditionally achieved with formaldehyde. Crosslinking performed directly in cells locks in the protein-DNA complexes, trapping these unstable and sometimes transient interactions. To lyse, extract and solubilize the crosslinked complexes, the ChIP assay kit includes the Thermo Scientific Chromatin Prep Module (also sold separately, Part No. 26158). Together, this complete ChIP assay kit provides a simple, reliable and convenient means for isolating chromatin-bound DNA and enriching samples for the proteins of interest with less than 15% contamination from other cellular compartments - without a Dounce homogenizer. |
| 特点： |
| - Simple and fast ChIP assay protocol (< 8 hours)- Highly efficient isolation and lysis of nuclei- Easy and reproducible enzymatic digestion- Low-background and high-binding capacity Protein A/G agarose resin- Highly specific positive controls included: RNA polymerase II antibody and GAPDH PCR primers- Fast and reproducible spin-column format- High-recovery DNA purification- ChIP-validated antibodies available |
| 数据： |
A431 lung carcinoma cells were cultured in DMEM containing 10% FBS for 24 hours. Following a 24 hour serum withdrawal, half of the cultures plated were treated with 100ng/mL EGF for 10 minutes. Crosslinking was achieved using a final concentration of 1% formaldehyde in the media for 10 minutes. ChIP assays were performed according to the manufacturers' protocols. Quantitive real-time PCR data was obtained with a Bio-Rad iQ5™ Thermocycler, Abgene Absolute SYBR Green Fluorescein QPCR master mix, and primers designed to amplify a region of the human MYC promoter proximal to the transcription start site. The Pierce Agarose ChIP Kit exhibited the highest signal to noise ratio. The Agarose ChIP Kit is effective for profiling multiple transcription factorsA431 lung carcinoma cells were cultured in DMEM containing 10% FBS for 24 hours. Following a 24 hour serum withdrawal, half of the cultures plated were treated with 100ng/mL EGF for 10 minutes. Crosslinking was achieved using a final concentration of 1% formaldehyde in the media for 10 minutes. ChIP assays were performed with the Pierce ChIP Kit to determine binding of phosphorylated-STAT3, acetylated-CBP, CBP, trimethyl histone H3, acetylated-histone H3, histone H3, and RNA polymerase II to the proximal MYC promoter. Primary antibody amounts were determined empirically. Quantitive real-time PCR data was obtained with a Bio-Rad iQ5™ Thermocycler, Abgene Absolute SYBR Green Fluorescein QPCR master mix, and primers designed to amplify a region of the human MYC promoter proximal to the transcription start site. |
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