Co-IP （免疫共沉淀，co-inmunoprecipitation）是经典的利用抗体从样品中捕获靶蛋白及其互作蛋白、复合体的一项技术，能够特异性富集所研究的目的蛋白。 由于过程中采用了非变性条件，保留了互作及复合体的细胞内状态。 相对于 酵母双杂交 、 pull down 等方法，其特色之一便是捕获的蛋白互作发生在所研究的特定组织细胞内，保存了互作蛋白及复合体的“内源”状态。 Co-IP实验原理： 以抗体和抗原之间的专一性作用为基础的用于研究蛋白质相互作用的经典方法，是确定 …

GFP-Trap is the benchmarking reagent for one-step immunoprecipitation (IP) of GFP-fusion proteins. The GFP-Trap consists of an anti-GFP Nanobody coupled to beads. The GFP-Trap provides a high level of IP performance. Next to IP, GFP-Trap can be applied in Co-IP, Co-IP/MS, on-bead assays, and ChIP/RIP analysis. What is a Nanobody?

Sep 26, 2013 · In this protocol we describe an advanced version of co-IP analysis that uses real-time, single-molecule fluorescence imaging as its detection scheme. Bait proteins are pulled down onto the...

Anti-GFP mouse monoclonal antibody 3E6 (A11120) is useful for immunoprecipitation, immunocytochemical localization, and immunosorbent assays (ELISA).

Jul 7, 2015 · By using live-cell imaging and biochemical analysis, we demonstrate that Myo1b is a downstream effector of EphB–ephrinB signaling controlling cell repulsion by regulating the redistribution of myosin II in actomyosin fibers and the formation of filopodia at the interface of ephrinB1 and EphB2 cells.

Jun 1, 2018 · To improve Co-IP results, a tag protein (such as myc, EGFP, and FLAG) can be fused to the protein of interest and express in the cell exogenously and perform Co-IP using a highly specific readily commercially available antibody against the tag protein.

Oct 12, 2023 · IP：即指免疫沉淀（immunoprecipitation），用相应的抗体来纯化富集目的蛋白，例如图中“IP: Flag”的意思是用Flag标签抗体磁珠去拉Flag，与Flag融合表达的蛋白为诱饵蛋白，此时与该融合蛋白相互作用的其他蛋白都会因此被沉淀下来。 IB：即指 免疫印迹 （immunoblotting）或 western blot （WB），用来检测目的蛋白的存在与否，例如图中“IB: Flag”是指用Flag抗体做WB检测，看样本中是否存在Flag标记的蛋白，IB：Myc的含义同理；值得注 …

Dec 1, 2022 · The vectors offer distinct benefits, including an improved multiple cloning site (MCS) with unique cutting sites for more restriction enzymes and a set of N- and C-terminal 3xMYC, 3xFLAG and EGFP epitope tags for visualization and purification of expressed proteins.

Co-immunoprecipitation (Co-IP) is one of the most widely used methods to identify novel proteins that associate with a protein of interest or to determine complex formation between known proteins. For this technique, a protein of interest is captured using a specific antibody.

如果蛋白本身在细胞内表达量就比较低，不好检测出来（本文中的PKN2在HCT116和SW480细胞中表达量较低），针对这种情况建议就做一个过表达Co-IP实验。 一般来讲，非内源性蛋白的相互作用，主要是通过过表达来实现，通常为简化实验、提高IP效率会让外源蛋白带上标签（本文中的flag和HA标签）。 在293 T细胞中设置了不同剂量 (0、3和6 μg) HA –标签的PKN2-WT（过表达PKN2），查看HA –PKN2-WT和 flag–DUSP6的共表达情况。