Scratch Wound Healing Assay
作者:Yanling Chen日期:03/05/2012,浏览量:105864,Q&A: 6
The scratch wound healing assay has been widely adapted and modified to study the effects of a variety of experimental conditions, for instance, gene knockdown or chemical exposure, on mammalian cell ...
Transwell Cell Migration Assay Using Human Breast Epithelial Cancer Cell
作者:Yanling Chen日期:02/20/2012,浏览量:62749,Q&A: 7
Transwell migration assays have been widely used for studying the motility of different types of cells including metastatic cancer cells. The assay is also useful in screens for compounds that act as ...
Analysis of Protein Stability by the Cycloheximide Chase Assay
Comparison of protein stability in eukaryotic cells has been achieved by cycloheximide, which is an inhibitor of protein biosynthesis due to its prevention in translational elongation. It is broadly ...
Measurement of Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) in Culture Cells for Assessment of the Energy Metabolism
作者:Birte Plitzko and Sandra Loesgen日期:05/20/2018,浏览量:39314,Q&A: 0
Mammalian cells generate ATP by mitochondrial (oxidative phosphorylation) and non-mitochondrial (glycolysis) metabolism. Cancer cells are known to reprogram their metabolism using different ...
BODIPY 493/503 Staining of Neutral Lipid Droplets for Microscopy and Quantification by Flow Cytometry
显微镜观察BODIPY 493/503 染色中性脂滴以及用流式细胞仪量化
作者:Bo Qiu and M. Celeste Simon日期:09/05/2016,浏览量:37940,Q&A: 1
Lipid droplets (LDs) are ubiquitous, dynamic organelles and function as a storage depot for neutral lipids, including triglycerides and cholesterol esters (Walther and Farese, 2012). The movement of ...
Sulforhodamine B (SRB) Assay in Cell Culture to Investigate Cell Proliferation
作者:Esteban A. Orellana and Andrea L. Kasinski日期:11/05/2016,浏览量:35749,Q&A: 0
The SRB assay has been used since its development in 1990 (Skehan et al., 1990) to inexpensively conduct various screening assays to investigate cytotoxicity in cell based studies (Vichai ...
In vitro Protein Ubiquitination Assays
作者:Qingzhen Zhao and Qi Xie日期:10/05/2013,浏览量:34575,Q&A: 1
Ubiquitin can be added to substrate protein as a protein tag by the concerted actions of ubiquitin activating enzyme (E1), ubiquitin conjugating enzyme (E2) and ubiquitin protein ligase (E3). At the ...
Infectious Focus Assays and Multiplicity of Infection (MOI) Calculations for Alpha-herpesviruses
作者:Anna Sloutskin and Ronald S. Goldstein日期:11/20/2014,浏览量:33005,Q&A: 0
Titration of viral stocks is a critical process before any experimental use of the virus. Here we describe an infectious focus assay for several alphaherpesviruses, a titration method for ...
Isolation of Mouse Tumor-Infiltrating Leukocytes by Percoll Gradient Centrifugation
作者:Ying Liu, Keqiang Chen, Chunyan Wang, Wanghua Gong, Teizo Yoshimura, Ji Ming Wang and Mingyong Liu日期:09/05/2013,浏览量:32929,Q&A: 3
A hallmark of cancer-associated inflammation is the infiltration of leukocytes into tumors, which is believed to be recruited by chemokines. Some infiltrating leukocytes such as macrophages often ...
Differentiation of THP1 Cells into Macrophages for Transwell Co-culture Assay with Melanoma Cells
作者:Michael Peter Smith, Helen Young, Adam Hurlstone and Claudia Wellbrock日期:11/05/2015,浏览量:30095,Q&A: 2
Understanding how immune cells such as macrophages interact with cancer cells is of increasing interest, as cancer treatments move towards combing both targeted- and immuno-therapies in new treatment ...
ImmunoPrecipitation of Nuclear Protein with Antibody Affinity Columns
作者:Saïda Dadi, Dominique Payet-Bornet and Pierre Ferrier日期:02/05/2013,浏览量:28416,Q&A: 1
Co-Immunoprecipitation (Co-IP) is the method used to pull down protein partners of a protein of interest using an antibody that specifically binds to this specific protein in order to test ...
Novel Object Recognition for Studying Memory in Mice
作者:Tzyy-Nan Huang and Yi-Ping Hsueh日期:10/05/2014,浏览量:25881,Q&A: 0
Memory tests are important indexes of the brain functions for rodents behavior assay. Many memory tasks require external forces (e.g. electric shocks) or intrinsic forces (e.g. ...
Targeted Gene Mutation in Rice Using a CRISPR-Cas9 System
采用CRISPR-Cas9 系统进行水稻基因定点突变
作者:Kabin Xie, Bastian Minkenberg and Yinong Yang日期:09/05/2014,浏览量:25299,Q&A: 0
RNA-guided genome editing (RGE) using bacterial type II cluster regularly interspaced short palindromic repeats (CRISPR)–associated nuclease (Cas) has emerged as a simple and versatile tool for ...
Hemagglutination Inhibition (HI) Assay of Influenza Viruses with Monoclonal Antibodies
Heamagglutination is inhibited when antibodies are present because antibodies to the influenza virus will prevent attachment of the virus to red blood cells. The highest dilution of antibody that ...
Microbial Mutagenicity Assay: Ames Test
作者:Urvashi Vijay, Sonal Gupta, Priyanka Mathur, Prashanth Suravajhala and Pradeep Bhatnagar日期:03/20/2018,浏览量:19813,Q&A: 2
The Microbial mutagenicity Ames test is a bacterial bioassay accomplished in vitro to evaluate the mutagenicity of various environmental carcinogens and toxins. While Ames test is used to ...
Cell Synchronization by Double Thymidine Block
作者:Guo Chen and Xingming Deng日期:09/05/2018,浏览量:18949,Q&A: 1
Cell synchronization is widely used in studying mechanisms involves in regulation of cell cycle progression. Through synchronization, cells at distinct cell cycle stage could be obtained. Thymidine ...
Polysome Fractionation to Analyze mRNA Distribution Profiles
作者:Amaresh C. Panda, Jennifer L. Martindale and Myriam Gorospe日期:02/05/2017,浏览量:17439,Q&A: 5
Eukaryotic cells adapt to changes in external or internal signals by precisely modulating the expression of specific gene products. The expression of protein-coding genes is controlled at the ...
Multiple Stepwise Gene Knockout Using CRISPR/Cas9 in Escherichia coli
作者:Enrico König, Francesca Zerbini, Ilaria Zanella, Davide Fraccascia and Guido Grandi日期:01/20/2018,浏览量:16546,Q&A: 1
With the recent implementation of the CRISPR/Cas9 technology as a standard tool for genome editing, laboratories all over the world are undergoing one of the biggest advancements in molecular biology ...
Murine Bronchoalveolar Lavage
作者:Fan Sun, Gutian Xiao and Zhaoxia Qu日期:05/20/2017,浏览量:16300,Q&A: 1
A basic Bronchoalveolar lavage (BAL) procedure in mouse is described here. Cells and fluids obtained from BAL can be analyzed by Hema3-staining, immunostaining, Fluorescence-activated cell sorting ...
Dot Blot Analysis of N6-methyladenosine RNA Modification Levels
作者:Lisha Shen, Zhe Liang and Hao Yu日期:01/05/2017,浏览量:15813,Q&A: 3
N6-methyladenosine (m6A) is the most prevalent internal modification of eukaryotic messenger RNA (mRNA). The total amount of m6A can be detected by several methods, ...
Muscle Histology Characterization Using H&E Staining and Muscle Fiber Type Classification Using Immunofluorescence Staining
作者:Chao Wang, Feng Yue and Shihuan Kuang日期:05/20/2017,浏览量:14844,Q&A: 1
Muscle function is determined by its structure and fiber type composition. Here we describe a protocol to examine muscle histology and myofiber types using hematoxylin and eosin (H&E) and ...
Robust Generation of Knock-in Cell Lines Using CRISPR-Cas9 and rAAV-assisted Repair Template Delivery
作者:Giel Vandemoortele, Delphine De Sutter and Sven Eyckerman日期:04/05/2017,浏览量:14666,Q&A: 0
The programmable Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated nuclease 9 (Cas9) technology revolutionized genome editing by providing an efficient way to cut the ...
Single-molecule Fluorescence in situ Hybridization (smFISH) for RNA Detection in Adherent Animal Cells
作者:Gal Haimovich and Jeffrey E. Gerst日期:11/05/2018,浏览量:14621,Q&A: 1
Transcription and RNA decay play critical roles in the process of gene expression and the ability to accurately measure cellular mRNA levels is essential for understanding this regulation. Here, we ...
Isothermal Titration Calorimetry: A Biophysical Method to Characterize the Interaction between Label-free Biomolecules in Solution
作者:Andrea Saponaro日期:08/05/2018,浏览量:13460,Q&A: 0
This protocol can be applied to analyze the direct interaction between a soluble protein and a target ligand molecule using Isothermal Titration Calorimetry (ITC, Malvern). ITC allows the biophysical ...
Quantification of Plant Cell Death by Electrolyte Leakage Assay
作者:Noriyuki Hatsugai and Fumiaki Katagiri日期:03/05/2018,浏览量:11701,Q&A: 1
We describe a protocol to measure the electrolyte leakage from plant tissues, resulting from loss of cell membrane integrity, which is a common definition of cell death. This simple protocol is ...
Adapting the Smart-seq2 Protocol for Robust Single Worm RNA-seq
Most nematodes are small worms that lack enough RNA for regular RNA-seq protocols without pooling hundred to thousand of individuals. We have adapted the Smart-seq2 protocol in order to sequence the ...
Targeted Nucleotide Substitution in Mammalian Cell by Target-AID
作者:Takayuki Arazoe, Keiji Nishida and Akihiko Kondo日期:06/05/2017,浏览量:8625,Q&A: 0
Programmable RNA-guided nucleases based on CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated protein) systems have been applied to various type of cells as ...
Analysis of the Mitochondrial Membrane Potential Using the Cationic JC-1 Dye as a Sensitive Fluorescent Probe
作者:Farzane Sivandzade, Aditya Bhalerao and Luca Cucullo日期:01/05/2019,浏览量:8205,Q&A: 0
In recent years, fluorescent dyes have been frequently used for monitoring mitochondrial membrane potential to evaluate mitochondrial viability and function. However, the reproducibility of the ...
Chromatin Immunoprecipitation Experiments from Whole Drosophila Embryos or Larval Imaginal Discs
作者:Vincent Loubiere, Anna Delest, Bernd Schuettengruber, Anne-Marie Martinez and Giacomo Cavalli日期:06/05/2017,浏览量:7859,Q&A: 1
Chromatin Immunoprecipitation coupled either to qPCR (qChIP) or high-throughput sequencing (ChIP-Seq) has been extensively used in the last decades to identify the DNA binding sites of transcription ...
Detection of mRNA by Whole Mount in situ Hybridization and DNA Extraction for Genotyping of Zebrafish Embryos
作者:Rachna Narayanan and Andrew C. Oates日期:03/20/2019,浏览量:5962,Q&A: 0
In situ hybridization is used to visualize the spatial distribution of gene transcripts in tissues and in embryos, providing important information about disease and development. Current ...
Evaluation of Anticancer activity of Silver Nanoparticles on the A549 Human Lung Carcinoma Cell Lines through Alamar Blue Assay
作者:Nikita Sharma, Geeta Arya, R. Mankamna Kumari, Nidhi Gupta and Surendra Nimesh日期:01/05/2019,浏览量:5751,Q&A: 0
Silver nanoparticles have been widely studied to possess antimicrobial as well as anticancer activity, and have found its applications in various fields including pharmaceutical industry, ...
Generation of Chemically Induced Liver Progenitors (CLiPs) from Rat Adult Hepatocytes
作者:Takeshi Katsuda, Kazunori Hosaka and Takahiro Ochiya日期:01/20/2018,浏览量:5561,Q&A: 0
Primary mature hepatocytes (MHs) or their progenitor cells are candidate cell sources for cell transplantation therapy in severe liver diseases. However, stable culture of these cells or generation ...
In vitro Osteoclastogenesis Assays Using Primary Mouse Bone Marrow Cells
作者:Xueqian Zhuang and Guohong Hu日期:06/05/2018,浏览量:5479,Q&A: 0
Osteoclasts are a group of bone-absorbing cells to degenerate bone matrix and play pivotal roles in bone growth and homeostasis. The unbalanced induction of osteoclast differentiation ...
Differentiation of Human Induced Pluripotent Stem Cells (iPSCs) into an Effective Model of Forebrain Neural Progenitor Cells and Mature Neurons
人诱导多能干细胞 (iPSCs) 分化为前脑神经祖细胞和成熟神经元的有效模型
Induced Pluripotent Stem Cells (iPSCs) are pluripotent stem cells that can be generated from somatic cells, and provide a way to model the development of neural tissues in vitro. One ...
Straight Channel Microfluidic Chips for the Study of Platelet Adhesion under Flow
作者:Alexander Dupuy, Lining Arnold Ju and Freda H Passam日期:03/20/2019,浏览量:5358,Q&A: 0
Microfluidic devices have become an integral method of cardiovascular research as they enable the study of shear force in biological processes, such as platelet function and thrombus formation. ...
Induced Germinal Center B Cell Culture System
作者:Kei Haniuda and Daisuke Kitamura日期:02/20/2019,浏览量:5228,Q&A: 0
The germinal center (GC) is the site where B cells undergo clonal expansion, affinity-based selection, and differentiation into memory B cells or plasma cells. It has been difficult to elucidate ...
Extraction and Analysis of Pan-metabolome Polar Metabolites by Ultra Performance Liquid Chromatography–Tandem Mass Spectrometry (UPLC-MS/MS)
超高效液相色谱 - 串联质谱法(UPLC-MS/MS)提取和分析泛代谢组极性代谢产物
作者:Dania M. Malik, Seth Rhoades and Aalim Weljie日期:02/05/2018,浏览量:4487,Q&A: 0
Modern triple quadrupole mass spectrometers provide the ability to detect and quantify a large number of metabolites using tandem mass spectrometry (MS/MS). Liquid chromatography (LC) is ...
In vitro Generation of CRISPR-Cas9 Complexes with Covalently Bound Repair Templates for Genome Editing in Mammalian Cells
The CRISPR-Cas9 system is a powerful genome-editing tool that promises application for gene editing therapies. The Cas9 nuclease is directed to the DNA by a programmable single guide (sg)RNA, and ...
Platelet Migration and Bacterial Trapping Assay under Flow
作者:Shuxia Fan, Michael Lorenz, Steffen Massberg and Florian Gaertner日期:09/20/2018,浏览量:3055,Q&A: 0
Blood platelets are critical for hemostasis and thrombosis, but also play diverse roles during immune responses. We have recently reported that platelets migrate at sites of infection in vitro ...
Unbiased and Tailored CRISPR/Cas gRNA Libraries by Synthesizing Covalently-closed-circular (3Cs) DNA
通过合成共价闭合环状(3Cs)DNA构建无偏和专用CRISPR / Cas gRNA文库
作者:Martin Wegner, Koraljka Husnjak and Manuel Kaulich日期:01/05/2020,浏览量:1758,Q&A: 0
Simplicity, efficiency and versatility of the CRISPR/Cas system greatly contributed to its rapid use in a broad range of fields. Applications of unbiased CRISPR/Cas screenings are increasing and thus ...