biological source
rabbit
antibody form
culture supernatant
clone
monoclonal
species reactivity
human
manufacturer/tradename
ChIPAb+, Upstate®
technique(s)
ChIP: suitable, immunoprecipitation (IP): suitable, western blot: suitable
isotype
IgG
NCBI accession no.
UniProt accession no.
shipped in
dry ice
Quality Level
相关类别
General description
ChIPAb+乙酰组蛋白H4(Lys5)套装包含乙酰组蛋白H4(Lys5)抗体、阴性对照上清、qPCR引物(用于扩增人GAPDH的166bp区)。乙酰组蛋白H4(Lys5)抗体和阴性对照的规格与ChIP反应次数成比例,可用于功能性验证乙酰组蛋白H4(Lys5)相关染色质的免疫沉淀。
Immunogen
Application
采用2 µL阴性对照上清液或2 µL抗乙酰组蛋白H4(Lys5)抗体和Magna ChIP® A试剂盒(货号17-610),对未处理HeLa细胞来源、超声处理的染色质(每次IP需1 X 106细胞当量)进行染色质免疫沉淀。
采用质控引物(用于扩增人GAPDH启动子)或引物(用于扩增HeLa细胞中转录失活的人β-珠蛋白启动子),经qPCR反应鉴定乙酰组蛋白H4(Lys5)相关DNA片段,验证免疫沉淀实验成功。
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。
实验详细信息请参阅EZ-Magna ChIP A(货号17-408)或EZ-ChIP(货号17-371)试剂盒方案。
蛋白质印迹分析:
代表性批次数据。
用1:1,000稀释的抗乙酰组蛋白H4(Lys5)抗体检测未处理或丁酸钠处理的HeLa细胞裂解液(分别为泳道1和2)。
用山羊抗兔抗体进行蛋白显色。(见图)。
蛋白质印迹分析和肽抑制:代表性批次数据。
电泳分离丁酸钠处理HeLa细胞的酸提取物,转印至PVDF膜,用1:1000稀释的抗乙酰组蛋白H4(Lys5)抗体检测。
为了验证抗体的特异性,用修饰的组蛋白H4肽预孵育血清:
泳道1:无多肽
泳道2:乙酰组蛋白H4(K5)(货号12-343)
泳道3:乙酰组蛋白H4(K8)(货号12-344)
泳道4:乙酰组蛋白H4(K12)(货号12-345)
泳道5:乙酰组蛋白H4(K16)(货号12-346)
泳道6:乙酰组蛋白H4(K5, 8, 12, 16)(货号12-353)
泳道7:组蛋白H4(货号12-347)
用HRP标记的驴抗兔IgG和化学发光检测系统对蛋白显色。(请参见图)。
组蛋白
表观遗传学&核功能
Biochem/physiol Actions
Packaging
Physical form
阴性对照上清液(兔):1瓶,50 μL培养物上清液,含0.05%叠氮化钠。储存在-20 °C。
对照引物。1瓶,75 μL,人GAPDH启动子特异性引物各5μM。在-20°C下储存。
对于:TAC TAG CGG TTT TAC GGG CG
REV: TCG AAC AGG AGG AGC AGA GAG CGA
Preparation Note
Analysis Note
包括阴性对照上清和人GAPDH特定引物。
采用2 µL阴性对照上清液或2 µL抗乙酰组蛋白H4(Lys5)抗体和Magna ChIP® A试剂盒(货号17-610),对来自未处理HeLa细胞的超声处理的染色质(每次IP需1 X 106细胞当量)进行染色质免疫沉淀。
使用针对人GAPDH启动子的ChIP质控引物经qPCR反应鉴定乙酰组蛋白H4(Lys5)相关DNA片段,验证免疫沉淀实验成功(见图)。
有关实验详细信息,请参阅EZ-Magna ChIPA(目录号17-408)或EZ-ChIP(目录号17-371)方案。
Legal Information
Disclaimer
存储类别
10 - Combustible liquids
法规信息
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.
Technical Guide to Chromatin Immunoprecipitation: Critical Factors for Success
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.
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