• Volume 11, 2021
  • Volume 10, 2020
  • 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
Image Credit: BioProtoc.2929


In vitro Enzymatic Assays of Histone Decrotonylation on Recombinant Histones Authors:  Rachel Fellows and Patrick Varga-Weisz, date: 07/20/2018, view: 4432, Q&A: 0
Class I histone deacetylases (HDACs) are efficient histone decrotonylases, broadening the enzymatic spectrum of these important (epi-)genome regulators and drug targets. Here, we describe an in vitro approach to assaying class I HDACs with different acyl-histone substrates, including crotonylated histones and expand this to examine the ...
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BMV Propagation, Extraction and Purification Using Chromatographic Methods Authors:  Aleksander Strugała, Paulina Bierwagen, Jakub Dalibor Rybka, Michał Giersig, Marek Figlerowicz and Anna Urbanowicz, date: 07/20/2018, view: 4182, Q&A: 0
Brome mosaic virus (BMV) is a well-known plant virus representing single-stranded RNA (ssRNA) positive-sense viruses. It has been widely used as a model in multiple studies concerning plant virus biology, epidemiology and the application of viral capsids in nanotechnology. Herein, we describe a method for BMV purification based on ion-exchange and ...
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Two Different Methods of Quantification of Oxidized Nicotinamide Adenine Dinucleotide (NAD+) and Reduced Nicotinamide Adenine Dinucleotide (NADH) Intracellular Levels: Enzymatic Coupled Cycling Assay and Ultra-performance Liquid Chromatography (UPLC)-Mass Spectrometry Authors:  Karina S. Kanamori, Guilherme C. de Oliveira, Maria Auxiliadora-Martins, Renee A. Schoon, Joel M. Reid and Eduardo N. Chini, date: 07/20/2018, view: 4885, Q&A: 0
Current studies on the age-related development of metabolic dysfunction and frailty are each day in more evidence. It is known, as aging progresses, nicotinamide adenine dinucleotide (NAD+) levels decrease in an expected physiological process. Recent studies have shown that a reduction in NAD+ is a key factor for the ...
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Measuring CD38 Hydrolase and Cyclase Activities: 1,N6-Ethenonicotinamide Adenine Dinucleotide (ε-NAD) and Nicotinamide Guanine Dinucleotide (NGD) Fluorescence-based Methods Authors:  Guilherme C. de Oliveira, Karina S. Kanamori, Maria Auxiliadora-Martins, Claudia C. S. Chini and Eduardo N. Chini, date: 07/20/2018, view: 5467, Q&A: 0
CD38 is a multifunctional enzyme involved in calcium signaling and Nicotinamide Adenine Dinucleotide (NAD+) metabolism. Through its major activity, the hydrolysis of NAD+, CD38 helps maintain the appropriate levels of this molecule for all NAD+-dependent metabolic processes to occur. Due to current advances and ...
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Cell Biology

Fluorophore-Based Mitochondrial Ca2+ Uptake Assay Author:  Charles B. Phillips, date: 07/20/2018, view: 4891, Q&A: 0
The physiological importance of mitochondrial calcium uptake, observed in processes such as ATP production, intracellular calcium signaling, and apoptosis, makes desirable a simple, straightforward way of investigating this event with unambiguous results. The following protocol uses a calcium-sensitive, membrane-impermeable fluorophore to monitor ...
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Developmental Biology

Muscle Function Assessment Using a Drosophila Larvae Crawling Assay Authors:  Yanina Post and Achim Paululat, date: 07/20/2018, view: 4450, Q&A: 0
Here we describe a simple method to measure larval muscle contraction and locomotion behavior. The method enables the user to acquire data, without the necessity of using expensive equipment (Rotstein et al., 2018). To measure contraction and locomotion behaviour, single larvae are positioned at the center of a humidified Petri dish. ...
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Measurement of TLR4 and CD14 Receptor Endocytosis Using Flow Cytometry Authors:  Michael S. Schappe and Bimal N. Desai, date: 07/20/2018, view: 6600, Q&A: 0
After recognizing extracellular bacterial lipopolysaccharide (LPS), the toll-like receptor 4 (TLR4)-CD14 signaling complex initiates two distinct signaling pathways–one from the plasma membrane and the other from the signaling endosomes (Kagan et al., 2008). Understanding the early stages of TLR4 signal transduction therefore requires a ...
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Single and Multiplexed Gene Editing in Ustilago maydis Using CRISPR-Cas9 Authors:  Mariana Schuster, Christine Trippel, Petra Happel, Daniel Lanver, Stefanie Reißmann and Regine Kahmann, date: 07/20/2018, view: 5639, Q&A: 0
The smut fungus Ustilago maydis is an established model organism for elucidating how biotrophic pathogens colonize plants and how gene families contribute to virulence. Here we describe a step by step protocol for the generation of CRISPR plasmids for single and multiplexed gene editing in U. maydis. Furthermore, we describe the ...
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Molecular Biology

α-Synuclein Aggregation Monitored by Thioflavin T Fluorescence Assay Authors:  Michael M. Wördehoff and Wolfgang Hoyer, date: 07/20/2018, view: 8251, Q&A: 1
Studying the aggregation of amyloid proteins like α-synuclein in vitro is a convenient and popular tool to gain kinetic insights into aggregation as well as to study factors (e.g., aggregation inhibitors) that influence it. These aggregation assays typically make use of the fluorescence dye Thioflavin T as a sensitive ...
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Ex vivo Whole-cell Recordings in Adult Drosophila Brain Authors:  Alexa J. Roemmich, Soleil S. Schutte and Diane K. O’Dowd, date: 07/20/2018, view: 4917, Q&A: 0
Cost-effective and efficient, the fruit fly (Drosophila melanogaster) has been used to make many key discoveries in the field of neuroscience and to model a number of neurological disorders. Great strides in understanding have been made using sophisticated molecular genetic tools and behavioral assays. Functional analysis of neural ...
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Embryonic Intravitreous Injection in Mouse Authors:  Liyuan Cui, Yupu Diao and Jiayi Zhang, date: 07/20/2018, view: 3383, Q&A: 0
Axons of retinal ganglion cells (RGCs) relay visual information from the retina to lateral geniculate nucleus (LGN) and superior colliculus (SC), which are two major image-forming visual nuclei. Wiring of these retinal projections completes before vision begins. However, there are few studies on retinal axons at embryonic stage due to technical ...
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Evaluating Working Memory on a T-maze in Male Rats Authors:  Ahmed M. Hussein, Mekite Bezu and Volker Korz, date: 07/20/2018, view: 7396, Q&A: 0
Working memory is short-term memory, so temporal improvement does not reflect the consolidation of a memory trace, rather the functionality of the underlying neuronal circuits and molecular signaling cascades. The administration of drugs–either one-time or through daily injection–can elucidate the underlying mechanisms. The T-maze is especially ...
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Assessing Experience-dependent Tuning of Song Preference in Fruit Flies (Drosophila melanogaster) Authors:  Xiaodong Li, Hiroshi Ishimoto and Azusa Kamikouchi, date: 07/20/2018, view: 4540, Q&A: 0
In songbirds and higher mammals, early auditory experience during childhood is critical to detect and discriminate sound patterns in adulthood. However, the neural and molecular nature of this acquired ability remains elusive. Here, we describe a new behavioral paradigm with Drosophila melanogaster to investigate how the auditory ...
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Plant Science

Extraction of RNA from Recalcitrant Tree Species Paulownia elongata Authors:  Niveditha Ramadoss and Chhandak Basu, date: 07/20/2018, view: 4380, Q&A: 0
Isolation of pure RNA is the basic requisite for most molecular biology work. Plants contain polyphenols and polysaccharides, which can interfere with isolation of pure RNA from them. Especially hardwood tree species like Paulownia elongata have surplus amount of RNA-binding alkaloids, proteins and secondary metabolites that can further ...
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High Resolution Melting Temperature Analysis to Identify CRISPR/Cas9 Mutants from Arabidopsis Authors:  Cynthia Denbow, Sonia Carole Ehivet and Sakiko Okumoto, date: 07/20/2018, view: 5865, Q&A: 0
CRISPR/Cas9 made targeted mutagenesis and genome editing possible for many plant species. One of the ways that the endonuclease is used for plant genetics is the creation of loss-of-function mutants, which typically result from erroneous DNA repair through non-homologous end joining (NHEJ) pathway. The majority of erroneous repair events results ...
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