Paper-based Invasion Assays for Quantifying Cellular Movement in Three-dimensional Tissue-like Structures [0.03%]
基于纸张的入侵试验用于定量三维组织样结构中的细胞运动
C Chad Lloyd,Matthew W Boyce,Matthew R Lockett
C Chad Lloyd
To elucidate the chemical and environmental conditions that promote invasion of cancer cells, an assay is needed in which the chemical landscape of a tumor-like environment can be experimentally manipulated and probed. The three-dimensional...
Marc Hafner,Mario Niepel,Kartik Subramanian et al.
Marc Hafner et al.
We developed a Python package to help in performing drug-response experiments at medium and high throughput and evaluating sensitivity metrics from the resulting data. In this article, we describe the steps involved in (1) generating files ...
Yulia A Bogdanova,Carsten Schultz,Vsevolod V Belousov
Yulia A Bogdanova
Described here is a localized H2 O2 generation-detection system consisting of a yeast D-amino acid oxidase (DAAO) and two spectrally distinct variants of biosensor, HyPer2 and HyPerRed based on circularly permutated yellow and red fluoresce...
Mario Niepel,Marc Hafner,Mirra Chung et al.
Mario Niepel et al.
Measuring the potencies of small-molecule drugs in cell lines is a critical aspect of preclinical pharmacology. Such experiments are also prototypical of high-throughput experiments in multi-well plates. The procedure is simple in principle...
Detecting Membrane Protein-protein Interactions Using the Mammalian Membrane Two-hybrid (MaMTH) Assay [0.03%]
用于检测膜蛋白-蛋白相互作用的哺乳动物膜两杂交(MaMTH)分析法
Punit Saraon,Ingrid Grozavu,Sang Hyun Lim et al.
Punit Saraon et al.
Protein-protein interactions (PPIs) play an integral role in numerous cellular processes. Membrane protein interactions, in particular, are critical in cellular responses to stresses and stimuli, with dysfunction of these PPIs (e.g., due to...
A General Non-Radioactive ATPase Assay for Chromatin Remodeling Complexes [0.03%]
一种用于染色质重塑复合物的通用无放射性ATP酶测定法
Benjamin Z Stanton,Courtney Hodges,Gerald R Crabtree et al.
Benjamin Z Stanton et al.
Chromatin remodeling complexes couple the energy released from ATP hydrolysis to facilitate transcription, recombination, and repair mechanisms essential for a wide variety of biologic responses. While recombinant expression of the regulato...
UbFluor: A Fluorescent Thioester to Monitor HECT E3 Ligase Catalysis [0.03%]
UbFluor：一种用于检测HECT E3连接酶催化的荧光硫酯
David T Krist,Peter K Foote,Alexander V Statsyuk
David T Krist
HECT E3 ubiquitin ligases (∼28 are known) are associated with many phenotypes in eukaryotes and are important drug targets. However, assays used to screen for small molecule inhibitors of HECT E3s are complex and require ATP, Ub, E1, E2, a...
Biochemical, Biophysical and Cellular Techniques to Study the Guanine Nucleotide Exchange Factor, GIV/Girdin [0.03%]
研究鸟嘌呤核苷酸交换因子GIV/Girdin的生化、生物物理和细胞技术
Pradipta Ghosh,Nicolas Aznar,Lee Swanson et al.
Pradipta Ghosh et al.
Canonical signal transduction via heterotrimeric G proteins is spatiotemporally restricted, i.e., triggered exclusively at the plasma membrane, only by agonist activation of G protein-coupled receptors via a finite process that is terminate...
Cyclic Immunofluorescence (CycIF), A Highly Multiplexed Method for Single-cell Imaging [0.03%]
单细胞成像的高通量方法循环免疫荧光(CycIF)
Jia-Ren Lin,Mohammad Fallahi-Sichani,Jia-Yun Chen et al.
Jia-Ren Lin et al.
Cyclic Immunofluorescence (CycIF) is a public-domain method for performing highly multiplexed immunofluorescence imaging using a conventional epifluorescence microscope. It uses simple reagents and existing antibodies to construct images wi...
Tracking the Activity of mTORC1 in Living Cells Using Genetically Encoded FRET-based Biosensor TORCAR [0.03%]
利用遗传编码的FRET型生物传感器TORCAR在活细胞中追踪mTORC1的活性
Xin Zhou,Simin Li,Jin Zhang
Xin Zhou
Mechanistic target of rapamycin complex 1 (mTORC1) is a highly conserved serine/threonine protein kinase that responds to multiple distinct signals (e.g., growth factors, amino acids, stress, and energy level) and coordinates cell growth an...