In vitro analysis of endoplasmic-reticulum-to-golgi transport in mammalian cells [0.03%]
哺乳动物细胞内质网向高尔基体转运的体外分析
B B Allan,W E Balch
B B Allan
A temperature-sensitive mutant of vesicular stomatitis G protein is used to follow the movement of that protein from the endoplasmic reticulum to transport vesicles to cis-Golgi and finally medial/trans-Golgi by assessing the maturation of ...
In vitro translation [0.03%]
体外翻译
R Jagus,B Joshi,S Miyamoto et al.
R Jagus et al.
The two most frequently used systems for in vitro translation are the rabbit reticulocyte system and the wheat germ extract. These systems are useful for mRNAs isolated from cells, tissues, and capped or uncapped mRNA produced in vitro by t...
N Rooke,J Underwood
N Rooke
Almost every eukaryotic pre-mRNA generated by RNA polymerase II transcription requires the removal of introns to create mRNA. The correct splicing of constitutive exons is thus critical for normal protein expression and function. Moreover, ...
D Duricka,K S Ullman
D Duricka
This unit describes a procedure for monitoring RNA export in Xenopus oocytes. The technique involves synthesizing labeled RNA in vitro and microinjecting the RNA into oocyte nuclei. Following incubation the oocytes are dissected into nuclea...
J Merlie Jr,R Heald
J Merlie Jr
The protocols in this unit describe the preparation of materials for an in vitro assay of mitotic spindle assembly in Xenopus egg extracts. Fluorochrome-labeled tubulin is used to visualize microtubule asters and spindles.
S Kornbluth,E K Evans
S Kornbluth
In the presence of a subcellular fraction enriched for mitochondria, after prolonged incubation the Xenopus egg extract can mimic biochemical aspects of apoptosis such as caspase activation, and DNA fragmentation. This unit describes prepar...
M Powers,E K Evans,J Yang et al.
M Powers et al.
In this unit, Xenopus eggs are isolated from hormonally primed female frogs, and then the extract is treated with cyclohexamide so it remains in interphase of the cell cycle. In the presence of sperm chromatin and ATP, membrane vesicles in ...
A Woods,J R Couchman
A Woods
Proteoglycans can be difficult molecules to isolate and analyze due to large mass, charge, and tendency to aggregate or form macromolecular complexes. This unit describes detailed methods for purification of matrix, cell surface, and cytosk...
S K Akiyama
S K Akiyama
This unit describes the purification of extracellular vitronectin from plasma by a series of affinity chromatography steps. First plasma is depleted of fibronectin and other heparin-binding proteins; the sample is then treated with urea to ...
Purification of fibronectin [0.03%]
纤粘蛋白的提纯
S K Akiyama
S K Akiyama
This unit describes the purification of the multifunctional adhesive glycoprotein fibronectin from plasma or of cell-derived fibronectin from cell surfaces and from conditioned medium. Fibronectin can be used in cell adhesion and migration ...