• Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011

Cancer Biology

In vivo Lineage-tracing Studies in a Cancer Stem Cell Population in Neuroblastoma Authors:  Danielle M. Hsu, Jason M. Shohet and Eugene S. Kim, date: 04/20/2014, view: 6401, Q&A: 0
Tumors are comprised of heterogeneous subpopulations that may exhibit differing capacity for differentiation, self-renewal, and tumorigenicity. In vivo lineage-tracing studies are a powerful tool for defining the role of tumor subpopulations in tumor growth and as targets for therapeutic agents. This protocol describes using a ...
More >>

Cell Biology

Purification and Fluorescent Labeling of Exosomes Authors:  Asuka Nanbo, Eri Kawanishi, Ryuji Yoshida and Hironori Yoshiyama, date: 04/20/2014, view: 13379, Q&A: 0
Exosomes are small membrane vesicles of endocytic origin secreted into the extracellular environment from a variety of different cells, and are thought to play important roles in intercellular communications. Here, we provide a useful protocol to purify the exosomes released from cell lines using sucrose gradient centrifugation. In this protocol, ...
More >>

Immunology

Quantification of T Cell Antigen-specific Memory Responses in Rhesus Macaques, Using Cytokine Flow Cytometry (CFC, also Known as ICS and ICCS): Analysis of Flow Data Authors:  Andrew W. Sylwester, Scott G. Hansen and Louis J. Picker, date: 04/20/2014, view: 10770, Q&A: 0
What was initially termed ‘CFC’ (Cytokine Flow Cytometry) is now more commonly known as ‘ICS’ (Intra Cellular Staining), or less commonly as ‘ICCS’ (Intra Cellular Cytokine Staining). The key innovations were use of an effective permeant (allowing intracellular staining), and a reagent to disrupt secretion (trapping cytokines, thereby enabling ...
More >>
Quantification of T Cell Antigen-specific Memory Responses in Rhesus Macaques, Using Cytokine Flow Cytometry (CFC, also Known as ICS and ICCS): from Assay Set-up to Data Acquisition Authors:  Andrew W. Sylwester, Scott G. Hansen and Louis J. Picker, date: 04/20/2014, view: 10546, Q&A: 0
What was initially termed ‘CFC’ (Cytokine Flow Cytometry’) is now more commonly known as ‘ICS’ (Intra Cellular Staining), or less commonly as ‘ICCS’ (Intra Cellular Cytokine Staining). The key innovations were use of an effective permeant (allowing intracellular staining), and a reagent to disrupt secretion (trapping cytokines, thereby enabling ...
More >>

Microbiology

Zonal Sedimentation Analysis on Sucrose Gradients Author:  Javier G. Magadán, date: 04/20/2014, view: 6163, Q&A: 0
Zonal sedimentation analysis on sucrose gradients allows estimation of the molecular size of an individual protein or a protein complex by centrifugation at a constant speed under nondenaturing conditions. This method is particularly suitable for globular proteins like the influenza A virus (IAV) protein hemagglutinin (HA). Here, I describe step ...
More >>
Radioactive Pulse-Chase Analysis and Immunoprecipitation Author:  Javier G. Magadán, date: 04/20/2014, view: 8533, Q&A: 0
Labeling of newly-synthesized polypeptides with radioactive amino acids followed by immunoprecipitation allows quantitative analysis of the fate of a given protein in a time-dependent manner. This biochemical approach is usually used to study a variety of processes, such as protein folding, co-translational modifications, intracellular transport, ...
More >>
Assay for Adherence of Vibrio cholerae to Eukaryotic Cell Lines Authors:  Amit K. Dey, Abha Bhagat and Rukhsana Chowdhury, date: 04/20/2014, view: 7367, Q&A: 0
Vibrio cholerae (V. cholerae) colonizes the intestinal epithelium and secretes cholera toxin (CT), a potent enterotoxin that causes severe fluid loss characteristic of the disease cholera. V. cholerae is a non-invasive Gram-negative bacterium that adheres to intestinal cells as well as a variety of different cell types. ...
More >>
Purification and Crystallization of Chloromuconolactone Dehalogenase ClcF from Rhodococcus opacus 1CP Authors:  Christian Roth, Janosch Alexander D. Gröning, Stefan Rudolf Kaschabek, Michael Schlömann and Norbert Sträter , date: 04/20/2014, view: 7252, Q&A: 0
The protocol describes the generation of variants of chloromuconolactone dehalogenase from Rhodococcus opacus (R. opacus) 1CP. ClcF is a multimeric protein, which catalyses the dechlorination of 5-chloromuconolactone to cis-dienelactone in the 3-chlorocatecholic acid degradation pathway. The protocol describes the ...
More >>
The BhbA Enzyme Assay Authors:  Kai Chen and Jiandong Jiang, date: 04/20/2014, view: 5309, Q&A: 0
Reductive dehalogenation has been found primarily in anaerobic communities and is originally thought to rarely occur in aerobes. A reductive dehalogenase (BhbA) was characterized from an aerobic strain of Comamonas sp. 7D-2, which was isolated from a bromoxynil octanoate-contaminated soil sample collected in Jiangsu, China. BhbA catalyzes ...
More >>

Plant Science

Protocol for the Preparation of Arabidopsis Meiotic Chromosome Spreads and Fluorescent in situ Hybridization Authors:  Pablo Bolaños-Villegas, Xiaohui Yang, Christopher A. Makaroff and Guang-Yuh Jauh, date: 04/20/2014, view: 10078, Q&A: 0
This protocol is a more detailed version of previous protocols (Yang et al., 2011; Bolaños-Villegas et al., 2013) developed for the examination of meiotic chromosome spreads. Meiotic chromosome spreads are useful to determine the presence of defects in chromosome pairing and segregation. The protocol also describes how to perform ...
More >>
Sandwich Enzyme-linked Immunosorbent Assay (ELISA) Analysis of Plant Cell Wall Glycan Connections Authors:  Valérie Cornuault and J. Paul Knox, date: 04/20/2014, view: 10698, Q&A: 0
Sandwich ELISA is a highly sensitive method that can be used to determine if two epitopes are part of the same macromolecule or supramolecular complex. In the case of plant cell wall glycans, it can reveal the existence of inter-polymers linkages, leading to better understanding of overall cell wall architectures. This development of a ...
More >>
Histochemical Detection of Superoxide and H2O2 Accumulation in Brassica juncea Seedlings Authors:  Deepak Kumar, Mohd Aslam Yusuf, Preeti Singh, Meryam Sardar and Neera Bhalla Sarin, date: 04/20/2014, view: 17482, Q&A: 2
Plant cells continually produce reactive oxygen species (ROS) as a by-product of aerobic metabolism. Increased production of ROS occurs under unfavorable conditions imposed by various abiotic and biotic factors. Accumulation of ROS is damaging to various cellular components and macromolecules including plasma membrane, nucleic acids, and proteins ...
More >>
We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.