ChIP如您所愿：快速+简单[新品推荐]

R&D Systems 细胞生物学研究工具
ExactaChIP™ 染色质免疫沉淀试剂盒

        R&D最新推出了一系列用于染色质免疫沉淀（ChIP）分析的ExactaChip试剂盒，包含FoxP3 (Catalog # ECP3240), beta-Catenin (Catalog # ECP1329), p53 (Catalog # ECP1355), Smad4 (Catalog # ECP2097), 和STAT5a/b (Catalog # ECP2168)。这一系列试剂盒可以简便快速地鉴定转录因子在基因组DNA上的结合位点。在交联的转录因子-DNA复合物免疫沉淀后，用普通的PCR和基因特异的引物来检测转录因子结合位点。每个ExactaChip试剂盒包含了一个免疫沉淀的抗体和一对已知转录因子结合DNA序列的引物作为阳性对照。研究者通过自己设计的引物，就可以发现转录因子结合的其他区域；通过试剂盒中提供的阳性对照引物，就可以检验ChIP反应的效果。ExactaChip试剂盒的更详细信息请访问：www.RnDSystems.com/go/ExactaChIP。

ChIP如您所愿：快速&简单

ü       抗体已验证过可用于染色质免疫沉淀

ü       快速——4-5小时内得到结果

ü       每个试剂盒配有一个阴性对照抗体

ü       结果稳定一致

ü       提供详尽易懂的操作步骤和问题解决指南

ü       每个试剂盒配有阳性对照引物对，更自信得到实验结果

 

实验原理     R&D 的ExactaChIP 染色质免疫沉淀试剂盒提供了一种快速、简单的方法来鉴定某个特定蛋白在基因组DNA上的靶定序列。通过甲醛交联固定蛋白-DNA复合物，再剪切染色质，通过这个目标蛋白的特异抗体对复合物进行免疫沉淀。随后，与蛋白质结合的DNA片段被纯化和通过PCR来进行扩增。   试剂盒组成 ü       分析用的特定一抗 ü       生物素化的对照抗体 ü       螯合树脂溶液 ü       对照引物对 ü       裂解液 ü       洗涤液 ü       稀释液   Detection of FoxP3 Genomic Targets by Chromatin Immunoprecipitation (IP). Human Jurkat T cells were stimulated with PMA and ionomycin, fixed, and lysed. FoxP3 binding to the IL-2 promoter was assessed using the FoxP3 ExactaChIP Chromatin IP Kit (Catalog # ECP3240). Lysed cells were incubated with biotinylated anti-human FoxP3 polyclonal antibody or biotinylated anti-goat IgG control, followed by MagCellect Streptavidin Ferrofluid (Catalog # MAG999). The DNA was purified and the IL-2 promoter was detected using standard PCR. M = DNA marker; Input = an aliquot of the total cell lysate DNA used for IP was added (+), or omitted (-) from the PCR reaction. .

Detection of GATA-5 Genomic Targets by Chromatin Immunoprecipitation. GATA-5 binding sites were assessed in HeLa cells using the GATA-5 ExactaChIP Chromatin IP Kit (Catalog # ECP2170). After fixation and lysis, cell lysates were incubated with anti-human GATA-5 polyclonal antibody followed by biotinylated anti-goat IgG polyclonal antibody (both provided in the kit) or with biotinylated anti-goat IgG polyclonal antibody alone. Immunocomplexes were captured using MagCellect Streptavidin Ferrofluid (Catalog # MAG999), and the DNA was purified using a chelating resin solution. The MUC4 promoter was detected by standard PCR using primers specific for MUC4 (provided in the kit). M=DNA Marker; Input = an aliquot of the total DNA used for immunoprecipitation was added to the PCR reaction