生物来源
rabbit
质量水平
克隆
polyclonal
种属反应性
human, mouse, vertebrates
制造商/商品名称
ChIPAb+
Upstate®
技术
ChIP: suitable
dot blot: suitable
western blot: suitable
NCBI登记号
UniProt登记号
运输
dry ice
基因信息
human ... H3F3B(3021)
相关类别
一般描述
ChIPAb+三甲基组蛋白H3(Lys9)套装包含抗三甲基组蛋白H3(Lys9)抗体、阴性对照抗体（纯化兔IgG）、qPCR引物（用于扩增人ZNF554基因3’端的117bp区域）。三甲基组蛋白H3(Lys9)抗体和阴性对照抗体的规格与ChIP反应次数成比例，可用于功能性验证三甲基组蛋白H3(Lys9)相关染色质的免疫沉淀。
免疫原
应用
采用4 μg的纯化抗体或正常兔IgG和Magna ChIP A试剂盒（货号17-610），对来自2x106个HeLa细胞的、超声制备的染色质进行染色质免疫沉淀。
使用针对人GAPDH启动子的ChIP引物和针对人MyoD启动子的引物，经qPCR鉴定三甲基组蛋白H3(Lys9)相关DNA片段，验证免疫沉淀成功。
详细实验方案请参阅 EZ-Magna ChIP A（货号17-408）或EZ-ChIP（货号17-371）试剂盒使用方法。
蛋白质印迹分析：
电泳分离重组组蛋白H3（泳道1）和HeLa细胞酸提取物（泳道2），转移至硝酸纤维素膜，用抗三甲基组蛋白H3(Lys9)抗体(1 μg/mL)检测。通过HRP标记山羊抗兔二抗和化学发光检测系统对蛋白显色（如图）。
Beadlyte组蛋白-肽特异性试验：
将0.5 μg/ml纯化抗二甲基组蛋白H3(Lys9)与具有下列修饰的组蛋白H3肽标记的微珠混合物共同孵育：
1. 三甲基-赖氨酸9
2. 二甲基-赖氨酸9
3. 单甲基-赖氨酸9
4.无修饰H3
过滤去除未结合抗体。将肽-抗体复合物与PE标记的抗兔二抗共同孵育。在Luminex 100仪器上读取荧光。绘制了中值荧光强度（MFI）。
生化/生理作用
外形
正常兔IgG。1小瓶，包含125 μg正常兔IgG，溶于125 μL储存液（含0.1%叠氮化钠）。储存于-20°C。
ChIP引物，ZNF554。1小瓶，包含75 μL ZNF554基因特定引物各5 μM。在-20°C下储存。
正向：CGG GGA AAA GCC CTA TAA AT
反向：TCC ACA TTC ACT GCA TTC GT
分析说明
包括阴性对照兔IgG抗体和对ZNF554特异性的对照引物。
采用4 µg正常兔IgG或抗三甲基组蛋白H3(Lys9)抗体和Magna ChIP A试剂盒（目录号17-610），对从3x106 NIH3T3 L1细胞超声制备的染色质进行染色质免疫沉淀。 使用ChIP引物ZNF554通过qPCR验证了三甲基组蛋白H3(Lys9)相关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 A ChIP（目录号17-408）或EZ-ChIP（目录号17-371）方案。
其他说明
ZNF554引物对，1瓶
法律信息
储存分类代码
10 - Combustible liquids
相关内容
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.
Technical Guide to Chromatin Immunoprecipitation: Critical Factors for Success
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 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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