ChIP如您所愿：快速+简单

实验原理
    R&D 的ExactaChIP染色质**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**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 ChromatinImmunoprecipitation (IP). Human Jurkat T cells were stimulated withPMA and ionomycin, fixed, and lysed. FoxP3 binding to the IL-2promoter was assessed using the FoxP3 ExactaChIP Chromatin IP Kit(Catalog # ECP3240). Lysed cells were incubated withbiotinylated anti-human FoxP3 polyclonal antibody or biotinylatedanti-goat IgG control, followed by MagCellect StreptavidinFerrofluid (Catalog # MAG999). The DNA was purified and the IL-2 promoterwas detected using standard PCR. M = DNA marker; Input = an aliquotof the total cell lysate DNA used for IP was added (+), or omitted(-) from the PCR reaction.

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Detection of GATA-5 Genomic Targets by ChromatinImmunoprecipitation. GATA-5 binding sites were assessed in HeLacells using the GATA-5 ExactaChIP Chromatin IP Kit (Catalog #ECP2170). After fixation and lysis, cell lysates were incubatedwith anti-human GATA-5 polyclonal antibody followed by biotinylatedanti-goat IgG polyclonal antibody (both provided in the kit) orwith biotinylated anti-goat IgG polyclonal antibody alone.Immunocomplexes were captured using MagCellect StreptavidinFerrofluid (Catalog # MAG999), and the DNA was purified using achelating resin solution. The MUC4 promoter was detected bystandard PCR using primers specific for MUC4 (provided in the kit).M=DNA Marker; Input = an aliquot of the total DNA used forimmunoprecipitation was added to the PCR reaction