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重组人巨噬细胞迁移抑制因子,His-标签(无动物源)(MIF)
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Chromatrap为Porvair Science公司旗下的产品。该品牌产品主要为ChIP相关试剂盒。Chromatrap® ChIP 测定技术比传统磁珠法的效率更高,原因是与蛋白质 A 或 G结合的固相多孔聚合物捕获染色质的效率更高。尽管同样是使用离心柱法,但其优势是琼脂糖珠或磁珠远不可相比的。
Chromatrap®为英国Porvair Science公司与斯旺西大学合作开发的产品。该品牌产品主要为ChIP相关试剂盒。Chromatrap® ChIP 测定技术比传统磁珠法的效率更高,原因是与蛋白质 A 或 G结合的固相多孔聚合物捕获染色质的效率更高,无须担心清洗过程中会意外丢失珠子。Chromatrap®技术获得的DNA Pull down 水平要比传统方法高出25倍,尤其适合于小型或低丰度生物样品。 ChIP是一种发展很快的研究技术,主要应用于确定细胞内DNA与蛋白质的相互作用,这一点对正确的基因调控尤为重要。ChIP在表观遗传学领域正被逐渐应用于药物研发中的分子机制研究、分层医学、诊断学以及将ChIP与第二代测序相结合的ChIP-seq技术中。 然而ChIP实验难度大,通常会遇到这样或那样的问题:如何确保较好的染色质质量、如何选择合适的片段化方式、超声处理和酶解方法有哪些优缺点、染色质片段化大小哪个范围合适、抗体的选择和设置又有哪些注意的地方? 当您在为遇到上述的问题食不知味、夜不能寐、山重水复疑无路时,不妨看看英国ChIP实验专家Chromatrap®提供的Top 10 tips锦囊,或许能柳暗花明又一村!
The use of high quality and specific ChIP validated antibodies is essential for the success of a ChIP assay. The antibody must recognise and bind to native protein that is bound to DNA. Antibodies from other applications do not always work well in ChIP.
Chromatrap® ChIP-validated Antibodies can now be supplied with the correct matching primer sets to cover all aspects of ChIP validation.
- Reduce time spent searching databases and designing forward and reverse primers
- Purchase superior quality ChIP-validated antibodies with their matching high-quality primers
- Have confidence that the primers are tested with the exact antibody and gene target
Many common epigenetic targets of interest are now available as combined antibody & primer sets from Chromatrap®.
In addition Chromatrap® supply forward and reverse primers on their own for common targets.
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文献和实验Active Motif ChIP 实验专家技术经验分享:如何判断 ChIP 成功与否?
对照转录因子)或 H3K4me3 的 ChIP(用于对照组蛋白修饰)。即使确定抗体可以用于做 ChIP,但对于目的细胞样本,如果之前没有做过,而且不确定目的蛋白结合的区域,一般也可以用别人报道过的对照细胞株作为对照,与目的细胞样本同时做 ChIP,确定 ChIP 是否成功。第二,整个实验是否真的 work?也就是如何排除假阳性?这就需要做阴性对照。阴性对照包括阴性对照抗体 IgG 和阴性对照区域(即阴性对照引物)。阴性对照 IgG 用于排除目的蛋白结合的 DNA 区域是抗体非特异性结合的,区域对照
Real-Time Primer Design for DNA Chips
is extensively used. The selection of the primers, which are immobilized on the DNA chip, requires a complex algorithm. Based on several parameters an optimized set of primers is automatically detected for a given gene sequence.This paper describes a parallel
Chromatin Immunoprecipitation (ChIP)
-deficient cells. Surprisingly, ~10% of the candidate sites that exhibited strong signals in the original ChIP-chip analysis, as well as in follow-up ChIP experiments performed with individual PCR primer pairs, yielded positive results of similar
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