High-Sensitivity ChIP试剂盒(ab185913)
Key features and details
- Sample type: Adherent cells, Suspension cells, Tissue
概述
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产品名称
High-Sensitivity ChIP试剂盒
参阅全部 ChIP Kit 试剂盒 -
样品类型
Tissue, Adherent cells, Suspension cells -
种属反应性
与反应: Mammals -
产品概述
High-Sensitivity ChIP Kit (ab185913) is a complete set of optimized reagents to carry out a successful chromatin immunoprecipitation procedure in a high throughput format starting from mammalian cells or tissues. The highly specific and sensitive kit is suitable for selective enrichment of a chromatin fraction containing specific DNA sequences using various mammalian cell/tissues. The optimized protocol and kit components reduce non-specific background ChIP levels to allow capture of low abundance protein/transcription factors and increased specific enrichment of target protein/DNA complexes. The target protein bound DNA prepared with the High-Sensitivity ChIP Kit can be used for various downstream applications including PCR (ChIP-PCR), microarrays (ChIP-on-chip), and sequencing (ChIP-seq).
Starting Materials
Starting materials can include various tissue or cell samples such as cells from flask or plate cultured cells, fresh and frozen tissues, etc. In general, the amount of cells and tissues for each reaction can be 2 x 103 to 1 x 106 and 0.5 mg to 50 mg, respectively. For optimal preparation, the input amount should be 1-2 x 105 cells or 10-20 mg tissues since the enrichment of target proteins to genome loci may vary. For the target proteins that are low abundance transcription factors, the input amount should be 5-6 x 105 cells or 50 to 60 mg tissues.
Primers
The GAPDH primers provided with the kit are for the human sequence. If using the kit with a difference species, GAPDH primers for that species will need to be acquired. -
说明
Protein-DNA interaction plays a critical role for cellular functions such as signal transduction, gene transcription, chromosome segregation, DNA replication and recombination, and epigenetic silencing. Identifying the genetic targets of DNA binding proteins and knowing the mechanisms of protein-DNA interaction is important for understanding cellular processes. Chromatin immunoprecipitation (ChIP) offers an advantageous tool for studying such protein-DNA interactions. It allows for the detection of a specific protein bound to a specific gene sequence in living cells using PCR (ChIP-PCR), microarrays (ChIP-chip), or sequencing (ChIP-seq). For example, measurement of the amount of methylated histone H3 at lysine 9 (meH3-K9) associated with a specific gene promoter region under various conditions can be achieved through a ChIP-PCR assay, while the recruitment of methylated H3-K9 to the promoters on a genome-wide scale can be detected by ChIP-on-chip or ChIP-sequencing. ChIP analysis requires that ChIPed DNA contains minimal background in order to reliably identify true TF-enriched regions. High background in ChIP is mainly caused ineffective wash buffers, insufficient cross-link reversal, inappropriate DNA fragment length, and residual RNA interference. To effectively capture TF/DNA complexes, which are often in low abundance, an ideal ChIP method requires having maximum sensitivity with minimized background levels. This method should also be able to enrich highly abundant protein/DNA complexes using a small amount of cells or tissues in a high throughput format. The High-Sensitivity ChIP Kit is designed to achieve these goals by maximizing sensitivity and minimizing non-specific background signals.
ChIP assay products and guides
Find more ChIP assay / chromatin immunoprecipitation resources and products, ChIP antibody products, and other ChIP assay kits and related reagents.
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经测试应用
适用于: ChIPmore details
性能
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存放说明
Please refer to protocols. -
组件 24 tests 48 tests 1000X Protease Inhibitor Cocktail 1 x 30µl 1 x 60µl 8-Well Assay Strips (with Frame) 3 units 6 units 8-Well Strip Caps 3 units 6 units Adhesive Covering Film 1 unit 2 units Antibody Buffer 1 x 3ml 1 x 6ml Anti-RNA Polymerase II 1 x 8µl 1 x 16µl Blocker Solution 1 x 2ml 1 x 4ml ChIP Buffer 1 x 6ml 1 x 12ml DNA Binding Solution 1 x 7ml 1 x 14ml DNA Elution Buffer 1 x 1ml 1 x 2ml DNA Release Buffer 1 x 8ml 1 x 16ml Enrichment Enhancer 1 x 55µl 1 x 110µl F-Collection Tube 30 units 50 units F-Spin Column 30 units 50 units GAPDH Primer - Forward (20 µM) 1 x 8µl 1 x 16µl GAPDH Primer - Reverse (20 µM) 1 x 8µl 1 x 16µl Lysis Buffer 1 x 14ml 1 x 28ml Non-Immune IgG (1 mg/ml) 1 x 10µl 1 x 20µl Proteinase K (10 mg/mL) 1 x 60µl 1 x 120µl Rnase A 1 x 30µl 1 x 60µl Wash Buffer 1 x 25ml 2 x 25ml -
研究领域
相关产品
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab185913于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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ChIP |
Use at an assay dependent concentration.
The GAPDH primers provided with the kit are for the human sequence. If using the kit with a different species, GAPDH primers for that species will need to be acquired. |
说明 |
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ChIP
Use at an assay dependent concentration. The GAPDH primers provided with the kit are for the human sequence. If using the kit with a different species, GAPDH primers for that species will need to be acquired. |
图片
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Sheared chromatin isolated from different numbers of MBD-231 cells was used for ChIP-qPCR analysis of RNA polymerase II enrichment in GAPDH promoters using ab185913 and a quantitative PCR Fast Kit.
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Sheared chromatin isolated from different numbers of MCF7 cells was used for ChIP-qPCR analysis of ER-a enrichment in TFF1 promoters using ab185913 and a quantitative PCR Fast Kit.
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Histone H3K18ac ChIP assay was carried out using ab185913.
ChIP-Seq reads align with the same peak sites as ENCODE data.
数据表及文件
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SDS download
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Datasheet download
文献 (30)
ab185913 被引用在 30 文献中.
- Zeng J et al. A Hypoxia-Induced SCFFBXL1 E3 Ligase Ubiquitinates and Degrades the MEN1 Tumor Suppressor to Promote Colorectal Cancer Tumorigenesis. Cancer Res Treat 54:525-540 (2022). PubMed: 34237211
- Shait Mohammed MR et al. The Histone H3K27me3 Demethylases KDM6A/B Resist Anoikis and Transcriptionally Regulate Stemness-Related Genes. Front Cell Dev Biol 10:780176 (2022). PubMed: 35186918
- Amato R et al. HIF-1-Dependent Induction of β3 Adrenoceptor: Evidence from the Mouse Retina. Cells 11:N/A (2022). PubMed: 35455951
- Zheng Y et al. Cancer-derived exosomal circ_0038138 enhances glycolysis, growth, and metastasis of gastric adenocarcinoma via the miR-198/EZH2 axis. Transl Oncol 25:101479 (2022). PubMed: 35987088
- Chavkin NW et al. Endothelial cell cycle state determines propensity for arterial-venous fate. Nat Commun 13:5891 (2022). PubMed: 36202789