{{'Search' | translate}}
 

CTS TM GlutaMAX TM-I Supplement

CTS TM GlutaMAX TM -I补充

Company: Thermo Fisher Scientific
Catalog#: A1286001
Bio-protocol()
Company-protocol()
Other protocol()

Cell-free Fluorescent Intra-Golgi Retrograde Vesicle Trafficking Assay
Author:
Date:
2017-11-20
[Abstract]  Intra-Golgi retrograde vesicle transport is used to traffic and sort resident Golgi enzymes to their appropriate cisternal locations. An assay was established to investigate the molecular details of vesicle targeting in a cell-free system. Stable cell lines were generated in which the trans-Golgi enzyme galactosyltransferase (GalT) was tagged with either CFP or YFP. Given that GalT is recycled to the cisterna where it is located at steady state, GalT-containing vesicles target GalT-containing cisternal membranes. Golgi membranes were therefore isolated from GalT-CFP expressing cells, while vesicles were prepared from GalT-YFP expressing ones. Incubating CFP-labelled Golgi with YFP-labelled vesicles in the presence of cytosol and an energy regeneration mixture at 37 °C produced a ... [摘要]  高尔基体内的逆行囊泡运输被用来运送和分类高尔基酶到适当的池内位置。建立了一个检测方法来研究无细胞系统中囊泡靶向的分子细节。生成了稳定的细胞系,其中反式 - 高尔基酶半乳糖基转移酶(GalT)用CFP或YFP标记。考虑到GalT被循环到稳定状态的小池中,含有GalT的囊泡将靶向含有GalT的池内膜。因此从表达GalT-CFP的细胞分离高尔基体膜,而从GalT-YFP表达细胞制备囊泡。在胞质溶胶和能量再生混合物的存在下,在37℃孵育CFP标记的高尔基体和YFP标记的囊泡,与在冰上孵育相比,在混合物的荧光成像后CFP-YFP共定位显着增加。该测定被验证需要能量,蛋白质和生理学重要的运输组分,如Rab GTP酶和保守寡聚体高尔基体系复合物。该测定法可用于调查影响高尔基体运输机器的生理和病理变化,特别是囊泡束缚。

【背景】细胞内囊泡靶向的分子机制对于解释糖基化稳态,神经递质释放,信号受体的调节和营养摄取等方面的解释是重要的(Ungar和Hughson,2003; ...

Neurosphere Co-culture Assay
Author:
Date:
2016-08-05
[Abstract]  The hippocampal niche is one of two areas in the brain where stem cells reside. In this neurogenic niche, stem cells can be found in close proximity to astrocytes and in contact with microvessels consisting of pericytes and endothelial cells. To study the regulatory interplay of this complex niche network in a simplified in vitro model, we established a co-culture system. We investigate the formation of neurosphere under different co-culture conditions by using primary niche cells. Here, we describe the isolation procedure for primary niche cells culture of astrocytes, endothelial cells and pericytes/smooth muscle cells from mouse brain. These niche cells are co-cultured (by hanging inserts) with freshly isolated stem and precursor cells from the adult hippocampus to study the ... [摘要]  真菌形态发生需要细胞壁的修饰和可塑性,这意味着其组分(包括壳多糖和葡聚糖)的合成和重塑。因此几丁质酶和葡聚糖酶活性对于细胞壁生物发生和细胞分裂是至关重要的。几丁质酶活性的定量可能有助于鉴定可能负面影响一些丝状真菌如产生细胞内和分泌型几丁质酶的尖孢镰刀菌的生长和形态发生的结构缺陷。基于它们对壳多糖底物的酶作用将壳聚糖分解酶分类。内酯酶被定义为催化几丁质链中内部点的随机裂解的酶。外切球蛋白酶催化乙酰壳寡糖或N-乙酰葡糖胺从几丁质的非还原端逐步释放,因此分别称为壳聚糖酶和β-N-乙酰氨基葡糖苷酶。在这里,我们描述了一种简单的方法来轻易地纯化几丁质酶,以便比较不同的F的内切几丁质酶活性和外切几丁质酶活性。尖孢镰孢菌株。该方案可适用于任何真菌物种。...

Comments