17-680
ChIPAb+ 单甲基组蛋白H3(Lys9) - ChIP免疫沉淀验证抗体和引物试剂套装
clone CMA306, from mouse, purified by using protein G
别名:
H3K9me1, Histone H3 (mono methyl K9)
相关类别
一般描述
ChIPAb+单甲基组蛋白H3(Lys9)套装包括抗单甲基组蛋白H3(Lys9)抗体，阴性对照抗体（纯化的小鼠IgG）和qPCR引物（可扩增人GAPDH基因编码区的213 bp）。 单甲基组蛋白H3(Lys9)和阴性对照抗体的规格与ChIP反应次数成比例，可用于功能上验证单甲基组蛋白H3(Lys9)相关染色质的沉淀。
免疫原
应用
采用2 µg的正常小鼠IgG或抗单甲基组蛋白H3(Lys9)抗体和Magna ChIP G（目录号17-611）试剂盒，对HeLa细胞来源的、超声处理的染色质（每次IP需1 X 106细胞）进行染色质免疫沉淀。 采用GAPDH编码区引物和GAPDH启动子区对照引物，经qPCR验证单甲基组蛋白H3(Lys9)相关DNA片段的成功免疫沉淀（请参见图）。数据表示为每个IP样本扩增子值相对于染色质输入(input)扩增子值的百分比(Percent Input)，ChIP样本如图所示。
请参阅EZ-Magna A ChIP（目录号17-408）或EZ-ChIP（目录号17-371）方案以获取实验详细信息。
蛋白质印迹分析：
前一批次的代表性数据。 将HeLa细胞酸提取物（泳道1）电泳分离，转印至PVDF膜，用抗单甲基组蛋白H3(Lys9)抗体(0.5 μg/mL)检测。
用HRP标记的山羊抗小鼠二抗（目录号AP124P）和化学发光检测系统对蛋白显色（如图）。
染色质生物学
表观遗传学&核功能
生化/生理作用
包装
外形
正常小鼠IgG。两个小瓶，包含25 μg纯化小鼠IgG，溶于25 μL储存缓冲液（含0.1%叠氮化钠）中。储存于-20°C。
ChIP引物，GAPDH编码区。一个小瓶，75 μL，人GAPDH编码区特定引物各5 μM。在-20°C下储存。
正向：GGC TCC CAC CTT TCT CAT CC
REV: GGC CAT CCA CAG TCT TCT GG
制备说明
分析说明
包括阴性对照小鼠IgG抗体和对人GAPDH编码区特异性的对照引物。
使用2 µg正常小鼠IgG或抗单甲基组蛋白H3(Lys9)抗体和Magna ChIP G（目录号17-611）试剂盒，对从HeLa细胞超声制备的染色质（每次IP需1 X 106细胞）进行染色质免疫沉淀。 使用ChIP引物（GAPDH编码区）通过qPCR验证了单甲基组蛋白H3(Lys9)作用DNA片段的成功免疫沉淀（请参见图）。
有关实验详细信息，请参阅EZ-Magna G ChIP（目录号17-409）或EZ-ChIP（目录号17-371）方案。
法律信息
免责声明
储存分类代码
10 - Combustible liquids
相关内容
Genome-wide mapping of DNA-protein interactions and epigenetic marks using chromatin immunoprecipitation combined with next generation sequencing (ChIP-Seq) is indispensable for many gene regulation studies. The Magna ChIPSeq Kit simplifies this technique, enabling the performance of ChIP-Seq by virtually any laboratory.
Epigenetics describes heritable changes in gene expression caused by non-genetic mechanisms. Epigenetic regulation allows a cell to vary its response based on its biological and environmental contexts. Epigenetic changes can effect transcriptional and post-transcriptional regulation via mechanisms such as histone modification, chromatin and nucleosome remodeling, DNA methylation, and small and non-coding RNA-mediated regulation. These mechanisms, in cooperation with transcription factors and other nucleic acid-binding proteins, regulate gene expression. Epigenetic mechanisms of gene regulation impacts diverse areas of research—from agriculture to human health. Common epigenetic assays such as chromatin immunoprecipitation (ChIP) and RNA immunoprecipitation (RIP) rely on high quality antibodies that recognize specific epigenetic modifications for accurate results. EMD Millipore offers over 100 ChIPAb+™ and RIPAb+™ validated antibody kits that are quality tested on ChIP/RIP assays and are conveniently provided with control qPCR primers and negative control antibodies to ensure first time ChIP/RIP success.
"Epigenetics describes heritable changes in gene expression caused by non-genetic mechanisms instead of by alterations in DNA sequence. These changes can be cell- or tissue-specific, and can be passed on to multiple generations. Epigenetic regulation enriches DNAbased information, allowing a cell to vary its response across diverse biological and environmental contexts. Although epigenetic mechanisms are primarily centered in the nucleus, these mechanisms can be induced by environmental signals such as hormones, nutrients, stress, and cellular damage, pointing to the involvement of cytoplasmic and extracellular factors in epigenetic regulation."
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