17-10086 EZ-Magna ChIP® A/G Chromatin Immunoprecipitation Kit免疫共沉淀

17-10086   Sigma-Aldrich  EZ-Magna ChIP® A/G Chromatin Immunoprecipitation Kit

Single day chromatin immunoprecipitation (ChIP) kit containing all necessary reagents to perform 22 individual chromatin immunoprecipitation (ChIP) reactions using magnetic A/G beads. Control primers included.

别名: Magnetic ChIP Kit

eCl@ss: 32161000

属性

质量水平

100

manufacturer/tradename

Magna ChIP®

technique(s)

immunoprecipitation (IP): suitable

运输

dry ice

说明

一般描述

Unlike standard ChIP protocols that can be laborious and time consuming, the Magna ChIP kit protocol can reduce the amount of time required to perform a ChIP experiment from three days to one. Additionally, the smaller Magna ChIP reaction volume increases the relative concentration of the antibody enabling the ChIP reaction to be performed with reduced amounts of both antibody and sheared chromatin. Finally because this kit uses a blend of protein A and protein G beads, a wider range of antibody isotypes can be used than A or G alone. This allows a wider variety of antibodies to be used and avoids the need to purchase separate kits for protein A and protein G based immunoprecipitation. Because Magna ChIP kits use paramagnetic beads they are compatible with automated high throughput platforms, thus allowing a large number of ChIP reactions to be carried out simultaneously. Features & Benefits:

• Magnetic bead-based rapid protocol allows performance of ChIP in as little as 1 day

• Blend of protein A+G bead blend allows ChIP using a broader range of antibodies than A or G alone

• Negative and positive control antibodies and control primer set to simplify validation of experimental procedure

• Includes spin columns to make DNA purification easier and more reliable - no more messy phenol-chloroform extractions

• Complete kit with all required reagents for reliable and reproducible results

• Compatible with ChIPAb+ validated antibody and primer sets

Chromatin Immunoprecipitation (ChIP) is an important technique allowing the analysis of in vivo interactions of proteins with genomic DNA. Any chromatin-associated or DNA binding protein can be analyzed with this technique, provided a good antibody to the protein exists. One can measure different proteins localized to a specific region of the genome, or the genome wide distribution of a specific protein. Another powerful application of this technique is to analyze changes in histone modifications that correlate with processes like transcription, mitosis or DNA repair.

Chromatin Immunoprecipitation (ChIP) is a powerful technique for mapping the in vivo distribution of proteins associated with chromosomal DNA. These proteins can be histone subunits and post-translational modifications or other chromatin associated proteins such as transcription factors, chromatin regulators, etc. Additionally, ChIP can be used to identify regions of the genome associated with these proteins, or conversely, to identify proteins associated with a particular region of the genome. ChIP methodology often involves protein-DNA and protein-protein cross-linking, fragmentation of the cross-linked chromatin, and subsequent immunoprecipitation of chromatin with an antibody specific to a target protein. The DNA fragments isolated in complex with the target protein can be identified by a variety of methods including PCR, DNA microarray and DNA sequencing. Standard or quantitative PCR can be performed to verify whether a particular DNA sequence (the gene or region of the genome) is associated with the protein of interest. The combination of ChIP and promoter or genomic tiling microarrays (ChIP-chip) allows genome-wide identification of DNA-binding sites for chromatin-associated proteins with precise resolution. Alternatively, high-throughput sequencing of libraries constructed from immunoprecipitated chromosomal DNA (ChIP-Seq) is a powerful alternative to ChIP-chip in mapping the protein-DNA interactions across mammalian genomes.

与费时费力的标准ChIP方法不同，Magna ChIP试剂盒方法可以将ChIP实验所需的时间从三天减少到一天。此外，较小的Magna ChIP反应体积增加了抗体的相对浓度，使得ChIP反应能够在抗体和剪切染色质数量减少的情况下进行。最后，由于该试剂盒使用蛋白质a和蛋白质G珠的混合物，因此可以使用比单独使用a或G更广泛的抗体同种型。这允许使用更多种类的抗体，并且避免了需要购买单独的试剂盒用于基于蛋白质a和蛋白质G的免疫沉淀。由于Magna ChIP方法使用顺磁珠，因此它们与自动化高通量平台兼容，从而允许同时进行大量芯片反应。

功能和优点：

基于磁珠的快速方法允许ChIP在一天内完成性能测试

蛋白质A+G珠混合物的混合物允许ChIP使用比单独使用A或G更广泛的抗体

阴性和阳性对照抗体以及对照引物组，以简化实验程序的验证

包括旋转柱，使DNA纯化更容易和更可靠-不再混乱酚氯仿提取

完整的试剂盒，配备所有必需的试剂，以获得可靠且可重复的结果

与ChIPAb+验证抗体和引物组兼容

染色质免疫沉淀（ChIP）是分析蛋白质与基因组DNA在体内相互作用的重要技术。任何染色质相关或DNA结合的蛋白质都可以用这种技术进行分析，只要存在一种良好的蛋白质抗体。人们可以测量定位于基因组特定区域的不同蛋白质，或特定蛋白质的全基因组分布。该技术的另一个强大应用是分析组蛋白修饰的变化，这些变化与转录、有丝分裂或DNA修复等过程相关。

染色质免疫沉淀（ChIP）是一种强有力的技术，用于绘制与染色体DNA相关的蛋白质在体内的分布图。这些蛋白质可以是组蛋白亚单位和翻译后修饰或其他染色质相关蛋白质，如转录因子、染色质调节器等。此外，ChIP可用于识别与这些蛋白质相关的基因组区域，或相反，识别与基因组特定区域相关的蛋白质。ChIP方法通常涉及蛋白质DNA和蛋白质-蛋白质交联、交联染色质的断裂，以及随后用特定于目标蛋白质的抗体对染色质进行免疫沉淀。与靶蛋白复合分离的DNA片段可以通过多种方法进行鉴定，包括PCR、DNA微阵列和DNA测序。可以进行标准或定量PCR来验证特定DNA序列（基因组的基因或区域）是否与感兴趣的蛋白质相关。ChIP与启动子或基因组拼接微阵列（ChIP-ChIP）的结合允许在全基因组范围内精确识别染色质相关蛋白质的DNA结合位点。此外，对免疫沉淀染色体DNA（ChIP-Seq）构建的文库进行高通量测序是在哺乳动物基因组中绘制蛋白质-DNA相互作用图谱的一种强有力的替代ChIP。

应用

Research Category
Epigenetics & Nuclear Function

Used to detect/quantify: Protein A+G

包装

Kit capacity: 22 chromatin immunoprecipitation assays

组分

Magnetic Protein A/G Beads

ChIP Dilution Buffer

Low Salt Wash Buffer

High Salt Wash Buffer

LiCl Wash Buffer

TE Buffer

Cell Lysis Buffer

Nuclear Lysis Buffer

ChIP Elution Buffer (w/o Proteinase K)

Proteinase K

RNase A

10X Glycine

10X PBS

Protease Inhibitor Cocktail II

Anti-RNA polymerase II, clone CTD4H8

Normal Mouse IgG

Control Primers

Spin Filters

Collection Tubes

Bind Reagent A

Wash Reagent B

Elution Reagent C

外形

Two boxes containing all necessary reagents to perform 22 individual chromatin immunoprecipitation (ChIP) reactions. Supplied buffers are sufficient to generate chromatin from up to five 15 cm plates of cultured cells, each plate providing up to 10 chromatin preparations (varies with cell and assay type).

储存及稳定性

Upon receipt, store components at the temperatures indicated on the labels. Kit components are stable for 1 year from date of shipment when stored as directed.

法律信息

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany

免责声明

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

安全信息

储存分类代码

10 - Combustible liquids

监管信息

危险化学品