• 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

Biochemistry

Phosphoinositides Coated Beads Binding Assay Authors:  Manuel Gálvez-Santisteban, Alejo E. Rodriguez-Fraticelli and Fernando Martin-Belmonte, date: 02/05/2014, view: 6356, Q&A: 0
The PIs coated beads assay or “PIP-Beads” developed by Echelon Biosciences (Salt Lake City, USA) is a quick assay to recognize which PIs are able to bind to a given protein domain, in a quantitative way. It is much faster and cheaper than liposomes and more reproducible than PIP-strip assays. The “PIP-Beads” assay is a biochemical assay that ...
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Cell Biology

Microsome Isolation from Tissue Authors:  Maria Bodero and Jose Francisco Abisambra, date: 02/05/2014, view: 7334, Q&A: 0
This protocol details the extraction of microsomes from frozen tissue in order to further examine the protein-protein interactions occurring within the endoplasmic reticulum. This protocol was adapted from Abisambra et al. (2013) with modifications made in order to optimize for subsequent use.
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Immunology

In vitro Biomineralization Assay Authors:  Kyunghee Lee, Minsuk Kwon and Daewon Jeong, date: 02/05/2014, view: 12079, Q&A: 1
Biomineralization in vertebrates has both physiological and pathological aspects. Physiological mineralization is essential for proper development and function of hard tissues, such as bone, teeth, and growth plate cartilage, but it does not occur in soft tissues. Pathological ectopic mineralization, in contrast, occurs in soft tissues, including ...
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Intracellular Staining for Phosphorylated STAT4 and STAT5 in Mouse Splenocytes Authors:  Ana Villegas-Mendez, J. Brian de Souza, Seen-Wai Lavelle, Emily Gwyer Findlay, Tovah N. Shaw, Christiaan J. Saris, Christopher A. Hunter, Eleanor M. Riley and Kevin N. Couper, date: 02/05/2014, view: 7415, Q&A: 0
The Stat (Signal Transducer and Activator of Transcription) family of proteins are critical signal transducers involved in fundamental cellular processes, including cell growth and differentiation, development, apoptosis, immune responses and inflammation. In here, we describe a simple and reproducible flow cytometry protocol to measure Stat ...
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Neuroscience

In situ Chemotaxis Assay in Caenorhabditis elegans (for the Study of Circadian Rhythms) Authors:  Martha Merrow and Maria Olmedo, date: 02/05/2014, view: 9131, Q&A: 0
Olfaction is a well-studied sensory mechanism in Caenorhabditis elegans (C. elegans). The nematodes respond to a wide range of chemicals by either attraction, repulsion or a mixture thereof (Bargmann et al., 1993). We have used olfaction to characterize behavioural and molecular circadian rhythms in C. elegans. ...
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Optical Clearing Using SeeDB Authors:  Meng-Tsen Ke, Satoshi Fujimoto and Takeshi Imai, date: 02/05/2014, view: 17612, Q&A: 1
We describe a water-based optical clearing agent, SeeDB (See Deep Brain), which clears fixed brain samples in a few days without quenching many types of fluorescent dyes, including fluorescent proteins and lipophilic neuronal tracers. SeeDB is a saturated solution of fructose (80.2% w/w) in water with 0.5% α-thioglycerol. In standard SeeDB optical ...
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Plant Science

Assays for Determination of Acetylesterase Activity and Specificity Using pNP-acetyl and Acetylated Polysaccharides as Substrates Authors:  Gennady Pogorelko and Olga A. Zabotina, date: 02/05/2014, view: 6645, Q&A: 0
The acetylesterases are hydrolytic enzymes which in plants cleave acetyl groups from acetylated cell wall components, primarily polysaccharides. To estimate acetylesterase activity in plant apoplast, two assays can be used. First assay is a direct measurement of the acetylesterase activity in protein extract using synthetic substrate, pNP-acetyl. ...
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Determination of Water Use Efficiency for Arabidopsis thaliana Authors:  Weronika Wituszynska and Stanisław Karpiński, date: 02/05/2014, view: 13424, Q&A: 1
Water use efficiency (WUE) is a quantitative measurement of how much biomass or yield is produced per amount of water used. It is an important physiological factor for agriculture, especially in areas with a limited accessibility of water. It is also crucial in a better understanding of drought tolerance and drought resistance. The most ...
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