RNA Whole-Mount In Situ Hybridization Proximity Ligation Assay (rISH-PLA), an Assay for Detecting RNA-Protein Complexes in Intact Cells [0.03%]
完整的细胞中用于检测RNA-蛋白质复合物的RNA原位杂交连接测定（rISH-PLA）
Ioannis M Roussis,Fiona A Myers,Garry P Scarlett
Ioannis M Roussis
Techniques for studying RNA-protein interactions have lagged behind those for DNA-protein interactions as a consequence of the complexities associated with working with RNA. This unit describes a method for the adaptation of the In Situ Hyb...
Proximity-Dependent Biotinylation for Identification of Interacting Proteins [0.03%]
邻近依赖性生物素标记技术在鉴定相互作用蛋白中的应用
Valerie Le Sage,Alessandro Cinti,Andrew J Mouland
Valerie Le Sage
Complex interaction networks orchestrate key cellular processes including but not limited to transcription, translation, metabolism, and cell signaling. Delineating these interactions will aid in deciphering the regulation and function of t...
LOVTRAP: A Versatile Method to Control Protein Function with Light [0.03%]
LOVTRAP：一种利用光控制蛋白质功能的灵活方法
Hui Wang,Klaus M Hahn
Hui Wang
We describe a detailed procedure for the use of LOVTRAP, an approach to reversibly sequester and release proteins from cellular membranes using light. In the application described here, proteins that act at the plasma membrane are held at m...
Isolation of Lipid Droplets from Cells by Density Gradient Centrifugation [0.03%]
通过密度梯度离心法从细胞中分离出脂滴
Dawn L Brasaemle,Nathan E Wolins
Dawn L Brasaemle
Lipid droplets are organelles found in most mammalian cells, as well as in various plant tissues and yeast. They are composed of a core of neutral lipids surrounded by a membrane monolayer of phospholipids and cholesterol in which specific ...
Identification of Protein Kinase Substrates by the Kinase-Interacting Substrate Screening (KISS) Approach [0.03%]
采用激酶底物筛选(KISS)法鉴定蛋白激酶的底物蛋白质
Mutsuki Amano,Tomoki Nishioka,Yoshimitsu Yura et al.
Mutsuki Amano et al.
Identifying the substrates of protein kinases to understand their modes of action has been undertaken by various approaches and remains an ongoing challenge. Phosphoproteomic technologies have accelerated the accumulation of data concerning...
Andrew D Doyle
Andrew D Doyle
Rat tail collagen solutions have been used as polymerizable in vitro three dimensional (3D) extracellular matrix (ECM) gels for single and collective cell migration assays as well as spheroid formation. Factors such as ECM concentration, pH...
Laura L Listenberger,Andrea M Studer,Deborah A Brown et al.
Laura L Listenberger et al.
Excess lipid is stored in intracellular organelles known as lipid droplets. This unit discusses techniques for the visualization of lipid droplets and associated proteins in cultured mammalian cells. Protocols for the detection of lipid dro...
Use of In Situ Proximity Ligation Assays for Systems Analysis of Signaling Pathways [0.03%]
原位邻近连接测定在信号转导途径系统分析中的应用
Tzu-Chi Chen,Chi-Ying F Huang
Tzu-Chi Chen
Understanding signaling pathway networks via protein-protein interactions (PPIs) at the cellular level is a significant task that has not yet been completed. Here, a systems approach that computationally infers interlinked pathways from num...
Preparation of Extracellular Matrices Produced by Cultured and Primary Fibroblasts [0.03%]
培养细胞和原代成纤维细胞产生的胞外基质的制备
Janusz Franco-Barraza,Dorothy A Beacham,Michael D Amatangelo et al.
Janusz Franco-Barraza et al.
Fibroblasts secrete and organize extracellular matrix (ECM), which provides structural support for their adhesion, migration, and tissue organization, besides regulating cellular functions such as growth and survival. Cell-to-matrix interac...
Immunoprecipitation [0.03%]
免疫沉淀技术
Juan S Bonifacino,David C Gershlick,Esteban C Dell&#x;Angelica
Juan S Bonifacino
Selective immunoprecipitation of proteins is a useful tool for characterizing proteins and protein-protein interactions. Clear step-by-step protocols are provided for preparing lysates of cells and yeast under a variety of conditions, for b...