• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

使用异源 Spike-ins 来标准化染色质免疫沉淀中技术变化的方案。

Protocol for using heterologous spike-ins to normalize for technical variation in chromatin immunoprecipitation.

机构信息

Metabolic Programming, School of Life Sciences Weihenstephan, ZIEL-Institute for Food & Health, Technische Universitaet Muenchen (TUM), 85354 Freising, Germany.

Institute for Diabetes and Obesity (IDO) & Institute for Diabetes and Cancer (IDC), Helmholtz Center Munich (HMGU) and German Center for Diabetes Research (DZD), 85764 Neuherberg (Munich), Germany.

出版信息

STAR Protoc. 2021 Jun 16;2(3):100609. doi: 10.1016/j.xpro.2021.100609. eCollection 2021 Sep 17.

DOI:10.1016/j.xpro.2021.100609
PMID:34189474
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8220248/
Abstract

Quantifying differential genome occupancy by chromatin immunoprecipitation (ChIP) remains challenging due to variation in chromatin fragmentation, immunoprecipitation efficiencies, and intertube variability. In this protocol, we add heterologous spike-ins from chromatin as an internal control to the mice chromatin before immunoprecipitation to normalize for technical variation in ChIP-qPCR or ChIP-seq. The choice of spike-in depends on the evolutionary conservation of the protein of interest and the antibody used. For complete details on the use and execution of this protocol, please refer to Greulich et al. (2021).

摘要

由于染色质片段化、免疫沉淀效率和管间变异性的差异，通过染色质免疫沉淀（ChIP）定量分析差异基因组占据仍然具有挑战性。在本方案中，我们在免疫沉淀前将来自 染色质的异源 Spike-ins 添加到小鼠染色质中，以 ChIP-qPCR 或 ChIP-seq 的技术变化为内参进行标准化。 Spike-in 的选择取决于感兴趣的蛋白质和所用抗体的进化保守性。有关此方案的使用和执行的完整详细信息，请参阅 Greulich 等人。（2021）。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/95cc1c3d199f/gr9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/6c9bb3972910/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/f37c052387a8/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/461bfd04301c/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/a74ca3d78993/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/5d46bde43429/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/b20d5797dddb/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/2eeab3749bf9/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/b65b5a5f9fd8/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/6f83df430b5a/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/7336e22a2955/fx2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/354f450fab53/fx3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/3e3968098ffc/fx4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/ba81cad0405d/fx5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/3392261f1873/fx6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/95cc1c3d199f/gr9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/6c9bb3972910/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/f37c052387a8/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/461bfd04301c/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/a74ca3d78993/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/5d46bde43429/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/b20d5797dddb/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/2eeab3749bf9/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/b65b5a5f9fd8/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/6f83df430b5a/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/7336e22a2955/fx2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/354f450fab53/fx3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/3e3968098ffc/fx4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/ba81cad0405d/fx5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/3392261f1873/fx6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc20/8220248/95cc1c3d199f/gr9.jpg

相似文献

1
Protocol for using heterologous spike-ins to normalize for technical variation in chromatin immunoprecipitation.使用异源 Spike-ins 来标准化染色质免疫沉淀中技术变化的方案。
STAR Protoc. 2021 Jun 16;2(3):100609. doi: 10.1016/j.xpro.2021.100609. eCollection 2021 Sep 17.
2
Protocol to apply spike-in ChIP-seq to capture massive histone acetylation in human cells.应用内掺 ChIP-seq 方法捕获人细胞中大量组蛋白乙酰化的方案。
STAR Protoc. 2021 Jul 17;2(3):100681. doi: 10.1016/j.xpro.2021.100681. eCollection 2021 Sep 17.
3
Quantitative Comparison of Multiple Chromatin Immunoprecipitation-Sequencing (ChIP-seq) Experiments with spikChIP.使用spikChIP对多个染色质免疫沉淀测序（ChIP-seq）实验进行定量比较。
Methods Mol Biol. 2023;2624:55-72. doi: 10.1007/978-1-0716-2962-8_5.
4
ChIPseq in Yeast Species: From Chromatin Immunoprecipitation to High-Throughput Sequencing and Bioinformatics Data Analyses.酵母物种中的染色质免疫沉淀测序：从染色质免疫沉淀到高通量测序及生物信息学数据分析
Methods Mol Biol. 2016;1361:185-202. doi: 10.1007/978-1-4939-3079-1_11.
5
Chromatin Preparation and Chromatin Immunoprecipitation from Drosophila Heads.从果蝇头部制备染色质和进行染色质免疫沉淀。
Methods Mol Biol. 2023;2655:19-30. doi: 10.1007/978-1-0716-3143-0_2.
6
Measuring Sister Chromatid Cohesion Protein Genome Occupancy in Drosophila melanogaster by ChIP-seq.通过染色质免疫沉淀测序（ChIP-seq）测量黑腹果蝇中姐妹染色单体黏连蛋白的基因组占有率
Methods Mol Biol. 2017;1515:125-139. doi: 10.1007/978-1-4939-6545-8_8.
7
Protocol for fractionation-assisted native ChIP (fanChIP) to capture protein-protein/DNA interactions on chromatin.分馏辅助天然染色质免疫沉淀（fanChIP）实验方案，用于捕获染色质上的蛋白质-蛋白质/DNA 相互作用。
STAR Protoc. 2021 Mar 23;2(2):100404. doi: 10.1016/j.xpro.2021.100404. eCollection 2021 Jun 18.
8
Chromatin-bound protein colocalization analysis using bedGraph2Cluster and PanChIP.使用 bedGraph2Cluster 和 PanChIP 进行染色质结合蛋白共定位分析。
STAR Protoc. 2023 Mar 17;4(1):101991. doi: 10.1016/j.xpro.2022.101991. Epub 2023 Jan 5.
9
A cell-sorting-based protocol for cell cycle small-scale ChIP sequencing.基于细胞分选的细胞周期小规模 ChIP 测序方案。
STAR Protoc. 2022 Mar 15;3(2):101243. doi: 10.1016/j.xpro.2022.101243. eCollection 2022 Jun 17.
10
Purification of mouse hepatic non-parenchymal cells or nuclei for use in ChIP-seq and other next-generation sequencing approaches.纯化小鼠肝脏非实质细胞或细胞核，用于染色质免疫沉淀测序（ChIP-seq）及其他新一代测序方法。
STAR Protoc. 2021 Mar 10;2(1):100363. doi: 10.1016/j.xpro.2021.100363. eCollection 2021 Mar 19.

引用本文的文献

1
Role of PRC2 in the stochastic expression of Aire target genes and development of mimetic cells in the thymus.PRC2在Aire靶基因的随机表达及胸腺中模拟细胞发育中的作用。
J Exp Med. 2025 Jul 7;222(7). doi: 10.1084/jem.20240817. Epub 2025 Apr 17.
2
Protocol to study the genomic profile of histone lactylation with CUT&RUN assay in tumor-associated macrophages.利用CUT&RUN分析研究肿瘤相关巨噬细胞中组蛋白乳酰化基因组图谱的方案。
STAR Protoc. 2025 Jun 20;6(2):103766. doi: 10.1016/j.xpro.2025.103766. Epub 2025 Apr 11.
3
ZNF143 is a transcriptional regulator of nuclear-encoded mitochondrial genes that acts independently of looping and CTCF.

本文引用的文献

1
The glucocorticoid receptor recruits the COMPASS complex to regulate inflammatory transcription at macrophage enhancers.糖皮质激素受体招募COMPASS复合物以调节巨噬细胞增强子处的炎症转录。
Cell Rep. 2021 Feb 9;34(6):108742. doi: 10.1016/j.celrep.2021.108742.
2
In Vivo ChIP-Seq of Nuclear Receptors: A Rough Guide to Transform Frozen Tissues into High-Confidence Genome-Wide Binding Profiles.核受体的体内染色质免疫沉淀测序：将冷冻组织转化为高可信度全基因组结合图谱的简要指南。
Methods Mol Biol. 2019;1966:39-70. doi: 10.1007/978-1-4939-9195-2_5.
3
CUT&Tag for efficient epigenomic profiling of small samples and single cells.
锌指蛋白143是核编码线粒体基因的转录调节因子，其作用独立于环化和CCCTC结合因子。
Mol Cell. 2025 Jan 2;85(1):24-41.e11. doi: 10.1016/j.molcel.2024.11.031. Epub 2024 Dec 20.
4
An Optimized High-Resolution Mapping Method for Glucocorticoid Receptor-DNA Binding in Mouse Primary Macrophages.一种优化的糖皮质激素受体-DNA 结合在小鼠原代巨噬细胞中高分辨率作图方法。
Methods Mol Biol. 2024;2846:91-107. doi: 10.1007/978-1-0716-4071-5_6.
5
Glucocorticoid Receptor-Dependent Binding Analysis Using Chromatin Immunoprecipitation and Quantitative Polymerase Chain Reaction.基于染色质免疫沉淀和实时定量聚合酶链反应的糖皮质激素受体依赖结合分析。
Methods Mol Biol. 2024;2846:17-34. doi: 10.1007/978-1-0716-4071-5_2.
6
Temporal evolution reveals bifurcated lineages in aggressive neuroendocrine small cell prostate cancer trans-differentiation.时间演变揭示侵袭性神经内分泌前列腺癌细胞转化的分支谱系。
Cancer Cell. 2023 Dec 11;41(12):2066-2082.e9. doi: 10.1016/j.ccell.2023.10.009. Epub 2023 Nov 22.
CUT&Tag 技术可高效地对小样本和单细胞进行表观基因组分析。
Nat Commun. 2019 Apr 29;10(1):1930. doi: 10.1038/s41467-019-09982-5.
4
Ensembl 2019.Ensembl 2019.
Nucleic Acids Res. 2019 Jan 8;47(D1):D745-D751. doi: 10.1093/nar/gky1113.
5
Purification of nanogram-range immunoprecipitated DNA in ChIP-seq application.ChIP-seq 应用中纳克级免疫沉淀 DNA 的纯化。
BMC Genomics. 2017 Dec 21;18(1):985. doi: 10.1186/s12864-017-4371-5.
6
The Encyclopedia of DNA elements (ENCODE): data portal update.《DNA 元件百科全书》（ENCODE）：数据门户更新。
Nucleic Acids Res. 2018 Jan 4;46(D1):D794-D801. doi: 10.1093/nar/gkx1081.
7
An efficient targeted nuclease strategy for high-resolution mapping of DNA binding sites.一种用于DNA结合位点高分辨率定位的高效靶向核酸酶策略。
Elife. 2017 Jan 16;6:e21856. doi: 10.7554/eLife.21856.
8
An Alternative Approach to ChIP-Seq Normalization Enables Detection of Genome-Wide Changes in Histone H3 Lysine 27 Trimethylation upon EZH2 Inhibition.一种用于ChIP-Seq标准化的替代方法能够检测EZH2抑制后全基因组组蛋白H3赖氨酸27三甲基化的变化。
PLoS One. 2016 Nov 22;11(11):e0166438. doi: 10.1371/journal.pone.0166438. eCollection 2016.
9
Differential peak calling of ChIP-seq signals with replicates with THOR.使用THOR对具有重复样本的ChIP-seq信号进行差异峰检测。
Nucleic Acids Res. 2016 Nov 16;44(20):e153. doi: 10.1093/nar/gkw680. Epub 2016 Aug 2.
10
Integrated genome browser: visual analytics platform for genomics.整合基因组浏览器：用于基因组学的可视化分析平台。
Bioinformatics. 2016 Jul 15;32(14):2089-95. doi: 10.1093/bioinformatics/btw069. Epub 2016 Mar 16.