Gene down-regulation with short hairpin RNAs and validation of specificity by inducible rescue in mammalian cells [0.03%]
哺乳动物细胞中用短发夹RNA使基因表达沉默及通过诱导型拯救验证其特异性
Hoi Tang Ma,Randy Y C Poon
Hoi Tang Ma
The principal problem with RNA interference (RNAi) experiments is off-target effects. The most vigorous demonstration of the specificity is the rescue of the RNAi effects with an RNAi-resistant target gene. By combining the expression of sh...
Analysis of DNA replication in Saccharomyces cerevisiae by two-dimensional and pulsed-field gel electrophoresis [0.03%]
用二维凝胶电泳和脉冲场凝胶电泳法分析Saccharomyces cerevisiae的DNA复制
Satoru Ide,Takehiko Kobayashi
Satoru Ide
Two methods to analyze DNA replication in budding yeast are described here. One is two-dimensional (2D) gel electrophoresis, which has been used to analyze replication intermediates in yeast. The other method is pulsed-field gel electrophor...
Simone Diermeier-Daucher,Gero Brockhoff
Simone Diermeier-Daucher
Dynamic proliferation assessment via flow cytometry is legitimately supposed to be the most powerful tool for recording cell cycle kinetics in-vitro. The preeminent feature is a single cell-based multi-informative analysis by temporal high-...
Carl E Creutz
Carl E Creutz
Adrenal medullary chromaffin granules (dense core secretory vesicles) have been a valuable model system for the study of the proteins and membrane components involved in the process of exocytosis. Because of the abundance of chromaffin gran...
Anatoli B Meriin,Yan Wang,Michael Y Sherman
Anatoli B Meriin
Upon permanent stresses and in various diseases, small protein aggregates may accumulate in cells and cause toxicity. A recently discovered protective system transports these aggregates to the centrosome location via microtubules, to form a...
Lee Ann Cohen,Julie G Donaldson
Lee Ann Cohen
This unit describes techniques and approaches that can be used to study the functions of the ADP-ribosylation factor (Arf) GTP-binding proteins in cells. There are six mammalian Arfs and many more Arf-like proteins (Arls), and these protein...
Vira V Artym,Kazue Matsumoto
Vira V Artym
The use of in vitro three-dimensional (3-D) collagen matrices to mimic an in vivo cellular environment has become increasingly popular and is broadening our understanding of cellular processes and cell-ECM interactions. To study cells in in...
Isolation of cytotoxic T cell and NK granules and purification of their effector proteins [0.03%]
细胞毒性T细胞和自然杀伤细胞颗粒的分离及其效应蛋白的纯化
Jerome Thiery,Michael Walch,Danielle K Jensen et al.
Jerome Thiery et al.
Killer lymphocytes induce apoptosis by the release of cytotoxic mediators from specialized secretory lysosomes, called cytotoxic granules, into the immunological synapse formed with a cell targeted for elimination. Methods are presented her...
Isolation of nucleoli [0.03%]
核仁的分离
Sabine Hacot,Yohann Coute,Stéphane Belin et al.
Sabine Hacot et al.
Nucleoli are now recognized as multi-functional nuclear domains involved in several fundamental cell processes such as ribosome biogenesis, regulation of the assembly of non-ribosomal ribonucleoprotein complexes, tRNA maturation, sequestrat...
Silencing of gene expression in cultured cells using small interfering RNAs [0.03%]
用小干扰RNA在培养细胞中使基因表达沉默
Kumi Sakurai,Pritsana Chomchan,John J Rossi
Kumi Sakurai
The discovery of RNA interference (RNAi) and related small RNA-mediated regulatory pathways has significantly altered the understanding of gene regulation in eukaryotic cells. In the RNAi pathway, small interfering RNAs (siRNAs) approximate...