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Image Credit: BioProtoc.3365


Subcellular Fractionation of Hela Cells for Lysosome Enrichment Using a Continuous Percoll-density Gradient 利用连续percoll密度梯度富集hela细胞溶酶体亚细胞组分 作者:Julian M. Carosi, Kathryn J. Hattersley, Yi Cui, Zhe Yang, Rohan D. Teasdale and Timothy J. Sargeant日期:09/20/2019,浏览量:5087,Q&A: 0
The enrichment of lysosomes is a useful way to study their structure and function. These dynamic vesicles can be enriched from cell cultures in a variety of ways including immunoprecipitation and fluorescence-activated organelle sorting. These methods are extremely precise but often require the transfection and expression of an affinity or ...
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A Cell Culture Model that Mimics Physiological Tissue Oxygenation Using Oxygen-permeable Membranes 一种利用透氧膜模拟生理组织氧合的细胞培养模式 作者:Chloe-Anne Martinez, Peter A. Cistulli and Kristina M. Cook日期:09/20/2019,浏览量:3739,Q&A: 0
Dissolved oxygen and its availability to cells in culture is an overlooked variable which can have significant consequences on experimental research outcomes, including reproducibility. Oxygen sensing pathways play key roles in cell growth and behavior and pericellular oxygen levels should be controlled when establishing in vitro models. ...
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SIRF: A Single-cell Assay for in situ Protein Interaction with Nascent DNA Replication Forks SIRF:一种蛋白与新生DNA复制叉原位相互作用的单细胞检测办法 作者:Sunetra Roy and Katharina Schlacher日期:09/20/2019,浏览量:3745,Q&A: 0
The duplication of DNA is a fundamental process that is required for the transfer of the genetic information from parent to daughter cells. Aberrant DNA replication processes are associated with diverse disease phenotypes, including developmental defects, ageing disorders, blood disorders such as Fanconi Anemia, increased inflammation and cancer. ...
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Characterization of Biological Motion Using Motion Sensing Superpixels 基于运动传感超像素的生物运动特性研究 作者:Felix Y. Zhou, Carlos Ruiz-Puig, Richard P. Owen, Michael J. White, Jens Rittscher and Xin Lu日期:09/20/2019,浏览量:3132,Q&A: 0
Precise spatiotemporal regulation is the foundation for the healthy development and maintenance of living organisms. All cells must correctly execute their function in the right place at the right time. Cellular motion is thus an important dynamic readout of signaling in key disease-relevant molecular pathways. However despite the rapid ...
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Supported Cell Membrane Sheets to Monitor Protein Assembly 蛋白质组装的细胞膜片监测 作者:Simon Erlendsson, Thor Seneca Thorsen and Kenneth Lindegaard Madsen日期:09/20/2019,浏览量:2995,Q&A: 0
Studying protein-protein and protein-lipid interactions in their native environment is highly desirable, yet, the heterogeneity and complexity of cellular systems limits the repertoire of experimental methods available. In cells, interactions are often taking place in confined microenvironments where factors such as avidity, hindered diffusion, ...
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Quantitation of TLR4 Internalization in Response to LPS in Thioglycollate Elicited Peritoneal Mouse Macrophages by Flow Cytometry 利用流式细胞术对巯基乙醇酸盐诱导的小鼠腹腔巨噬细胞中LPS-TLR4复合物内化的定量分析 作者:Katharina Richard, Stefanie N. Vogel and Darren J. Perkins日期:09/20/2019,浏览量:2483,Q&A: 0
Internalization of cell surface Toll-like Receptor 4 (TLR4) is a convenient read-out to measure LPS dependent activation of the TRIF adaptor pathway. We here provide a protocol to quantify the LPS dependent internalization of TLR4 using thioglycollate-elicited peritoneal macrophages by flow cytometry.
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Conjugation of Fab’ Fragments with Fluorescent Dyes for Single-molecule Tracking on Live Cells Fab片段结合荧光染料用于活细胞单分子示踪 作者:I-Ting Teng, Xiangning Bu and Inhee Chung日期:09/20/2019,浏览量:3919,Q&A: 0
Our understanding of the regulation and functions of cell-surface proteins has progressed rapidly with the advent of advanced optical imaging techniques. In particular, single-molecule tracking (SMT) using bright fluorophores conjugated to antibodies and wide-field microscopy methods such as total internal reflection fluorescence microscopy have ...
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Immunohistochemical Staining of CD8α in Diabetic Mouse Kidney 糖尿病小鼠肾脏CD8α的免疫组化染色 作者:Ming-Zhi Zhang and Raymond C. Harris日期:09/20/2019,浏览量:1704,Q&A: 0
Immune cell infiltration, particularly cytotoxic CD8α lymphocyte infiltration, plays an important role in development of diabetic nephropathy. Although CD8α infiltration can be evaluated by its production of cytokines, its localization in the kidney is of particular importance. The current protocol describes CD8α immunostaining using a Vectastain ...
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Production of Quantum Dots-containing Influenza Virus Particles for Studying Viral Uncoating Processes 量子点标记的流感病毒样颗粒用于病毒脱壳过程的研究 作者:Chong Qin, Xiaowei Zhang and Zongqiang Cui日期:09/20/2019,浏览量:3335,Q&A: 0
The genome of influenza A virus (IAV) comprises eight pinlike genomic segments called vRNPs enclosed in viral capsid. During infection, uncoating is the key step for viral replication and represents an antiviral therapeutic target, but it is difficult to observe the transient and dynamic event in detail. Here, we report a protocol for production ...
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Cryo-transmission Electron Microscopy of Outer-inner Membrane Vesicles Naturally Secreted by Gram-negative Pathogenic Bacteria 革兰阴性致病菌自然分泌的外-内膜囊泡低温透射电子显微镜观察 作者:Lidia Delgado, Nicolás Baeza, Carla Pérez-Cruz, Carmen López-Iglesias and Elena Mercadé日期:09/20/2019,浏览量:3228,Q&A: 0
A protocol was developed to visualize and analyze the structure of membrane vesicles (MVs) from Gram-negative bacteria. It is now accepted that these micrometric spherical vesicles are commonly produced by cells from all three domains of life, so the protocol could be useful in the study of vesicles produced by eukaryotes and archaea as well as ...
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A PhoA-STII Based Method for Efficient Extracellular Secretion and Purification of Fab from Escherichia coli 一种基于phoa stii技术的 大肠杆菌中高效分泌表达的fab片段纯化 作者:Ziyan Wang, Yang Gao, Manyu Luo, Cedric Cagliero, Hua Jiang, Yueqing Xie, Jianwei Zhu and Huili Lu日期:09/20/2019,浏览量:3836,Q&A: 0
In comparison with full-length IgGs, antigen binding fragments (Fabs) are smaller in size and do not require the complexed post-translational modification. Therefore, Fab can be cost-effectively produced using an Escherichia coli (E. coli) expression system. However, the disulfide-bonds containing exogenous protein, including ...
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In situ Hybridization of Plant-parasitic Nematode Globodera pallida Juveniles to Detect Gene Expression 原位杂交检测马铃薯白胞囊线虫的基因表达 作者:Joanna Kud, Nejra Solo, Allan Caplan, Joseph C. Kuhl, Louise-Marie Dandurand and Fangming Xiao日期:09/20/2019,浏览量:2701,Q&A: 0
In this study, we describe a standard whole mount in situ hybridization method which is used to determine the spatial-temporal expression pattern of genes from Globodera spp. Unlike more invasive radioactive labeling approaches, this technique is based on a safe, highly specific enzyme-linked immunoassay where a Digoxigenin ...
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Using indCAPS to Detect CRISPR/Cas9 Induced Mutations 利用indCAPS检测CRISPR/Cas9诱导的突变 作者:Charles Hodgens, Zachary L. Nimchuk and Joseph J. Kieber日期:09/20/2019,浏览量:2278,Q&A: 0
Cleaved amplified polymorphic sequences (CAPS) assays are useful tools for detecting small mutations such as single nucleotide polymorphisms (SNPs) or insertion/deletions (indels) present in an amplified DNA fragment. A mutation that disrupts or creates a restriction site will prevent cleavage by a restriction enzyme, allowing discrimination of ...
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Sciatic Nerve Cut and Repair Using Fibrin Glue in Adult Mice 纤维蛋白胶在成年小鼠坐骨神经损伤修复中的应用 作者:Erica T. Akhter, Travis M. Rotterman, Arthur W. English and Francisco J. Alvarez日期:09/20/2019,浏览量:2594,Q&A: 0
Peripheral nerve injury (PNI) is an excellent model for studying neural responses to injury and elucidating the mechanisms that can facilitate axon regeneration. As such, several animal models have been employed to study regenerative mechanisms after PNI, including Aplysia, zebrafish, rabbits, cats and rodents. This protocol describes how to ...
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Explant Culture of the Embryonic Mouse Spinal Cord and Gene Transfer by ex vivo Electroporation 小鼠胚胎脊髓的组织块培养和体外电穿孔介导的基因转移 作者:Mariko Kinoshita-Kawada, Hiroshi Hasegawa, Tsunaki Hongu, Shigeru Yanagi, Yasunori Kanaho, Ichiro Masai, Takayasu Mishima, Xiaoping Chen, Yoshio Tsuboi, Yi Rao, Junichi Yuasa-Kawada and Jane Y. Wu日期:09/20/2019,浏览量:3245,Q&A: 0
Developing axons change responsiveness to guidance cues during the journey to synapse with target cells. Axon crossing at the ventral midline serves as a model for studying how axons accomplish such a switch in their response. Although primary neuron culture has been a versatile technique for elucidating various developmental mechanisms, many in ...
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Simple Protocol for Distinguishing Drug-induced Effects on Spatial Memory Acquisition, Consolidation and Retrieval in Mice Using the Morris Water Maze 一种利用Morris水迷宫分辨药物诱导对于小鼠空间记忆获取、巩固和恢复学习行为影响的简单方案 作者:Ariel K. Frame, Asad Lone, Richard A. Harris and Robert C. Cumming日期:09/20/2019,浏览量:3742,Q&A: 0
The Morris water maze (MWM) is one of the most commonly used tests for assessing spatial learning and memory in mice. While the MWM is highly amenable to testing the effects of memory modifying drugs, most studies do not consider the timing or duration of drug exposure when conducting the MWM assay; factors that can strongly influence the effect ...
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