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Whatman Puradisc注射器过滤器

Company: Cytiva
Catalog#: 10462200
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Optogenetic Tuning of Ligand Binding to The Human T cell Receptor Using The opto-ligand-TCR System
[Abstract]  T cells are one major cell type of the immune system that use their T cell antigen receptor (TCR) to bind and respond to foreign molecules derived from pathogens. The ligand-TCR interaction half-lives determine stimulation outcome. Until recently, scientists relied on mutating either the TCR or its ligands to investigate how varying TCR-ligand interaction durations impacted on T cell activation. Our newly created opto-ligand-TCR system allowed us to precisely and reversibly control ligand binding to the TCR by light illumination. This system uses phytochrome B (PhyB) tetramers as a light-regulated TCR ligand. PhyB can be photoconverted between a binding (ON) and non-binding (OFF) conformation by 660 nm and 740 nm light illumination, respectively. PhyB ON is able to bind to a synthetic ... [摘要]  [摘要] T细胞是免疫系统的一种主要细胞类型,利用其T细胞抗原受体(TCR)结合并响应源自病原体的外来分子。配体-TCR相互作用的半衰期决定了刺激的结果。直到最近,科学家还是依靠突变TCR或其配体来研究变化的TCR-配体相互作用持续时间如何影响T细胞活化。我们新创建的光配体-TCR系统使我们能够通过光照精确且可逆地控制配体与TCR的结合。该系统使用植物色素B(PhyB )四聚体作为光调节的TCR配体。PHYB 可光转化的结合(ON)之间,并且通过分别为660nm和740nm的光照射,非结合性(OFF)构象。PhyB ON能够结合通过将PhyB 相互作用因子(PIF)融合到TCRβ链而产生的合成TCR 。将PhyB 切换为OFF构象会破坏这种相互作用。PhyB 四聚体与PIF-TCR 足够长的结合导致T细胞活化(通过钙内流测量)。在这里,我们描述了有关如何为我们的光配体-TCR系统生成四聚体配体,如何通过流式细胞仪测量配体-TCR结合以及如何通过钙内流量化T细胞活化的协议。

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Secretion Assay in Shigella flexneri
[Abstract]  Shigella flexneri (S. flexneri) is a Gram-negative bacterium that causes gastroenteritis and shigellosis in humans. In order to establish and maintain an infection, S. flexneri utilises a type three secretion system (T3SS) to deliver virulence factors called effector proteins into the cytoplasm of host cells, facilitating e.g. uptake into the host cell and escape from the endosome. Secretion through the T3SS is tightly regulated and is usually triggered by host-cell contact, but can also be artificially stimulated in vitro. In this assay, the dye Congo red is used to induce T3SS-dependent secretion of S. flexneri (Parsot et al., 1995) and secreted proteins are concentrated from the culture supernatant by precipitation with ... [摘要]  志贺氏菌(Shigella flexneri)( S.flexneri )是一种革兰氏阴性细菌,其在人类中引起胃肠炎和志贺氏菌病。为了建立和维持感染, flexneri利用三型分泌系统(T3SS)将称为效应子蛋白的毒力因子递送到宿主细胞的细胞质中,促进例如摄取到宿主细胞中并从内体中逃逸。通过T3SS的分泌是严格调节的,通常由宿主细胞接触引发,但也可以在体外人工刺激。在该测定中,染料刚果红用于诱导T3的依赖于T3SS的分泌。 (Parsot等人,1995),通过用三氯乙酸沉淀从培养物上清液中浓缩分泌的蛋白质。本文提供的测定可以容易地适应于利用T3SS(例如鼠伤寒沙门氏菌)的其他细菌的分泌分析,当在37℃下生长时,其组成性分泌(Collazo等, (Heesemann等人,1995; Pegues等人,1995)或Yersinia的致病物种,其中可通过钙剥夺诱导分泌(Heesemann等人,/em,1986; Forsberg等人,1987)。