• 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

Protein Extraction from Drosophila Embryos and Ovaries Authors:  Pedro Prudêncio and Leonardo G. Guilgur, date: 05/05/2015, view: 11833, Q&A: 0
Here we provide the description of protocols to efficiently obtain protein extracts from embryos and ovaries of Drosophila melanogaster. These protocols are routinely applied in our laboratory and are based on two techniques: either embryos or ovaries are homogenized using a pestle and then the soluble proteins separated by ...
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Microbiology

Quantification of HIV RNA and Human Herpesvirus DNA in Seminal Plasma Authors:  Milenka V. Vargas-Meneses, Marta Massanella, Caroline C. Ignacio and Sara Gianella, date: 05/05/2015, view: 5865, Q&A: 0
Multiple viruses can co-infect the genital tract, modifying the immunologic and virologic milieu and possibly playing a role in viral transmission and pathogenesis. The aim of our studies has been to understand the complex relationships between HIV-1 RNA, and multiple human herpesviruses known to frequently replicate in the genital tract of ...
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Measurement of Chlorophyll a and Carotenoids Concentration in Cyanobacteria Authors:  Tomáš Zavřel, Maria A. Sinetova and Jan Červený, date: 05/05/2015, view: 19431, Q&A: 1
This is a protocol for precise measurement of chlorophyll a and total carotenoid concentrations in cyanobacteria cells. Cellular chlorophyll concentration is one of the central physiological parameters, routinely followed in many research areas ranging from stress physiology to biotechnology. Carotenoids concentration is often related to ...
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Molecular Biology

FLP/FRT Induction of Mitotic Recombination in Drosophila Germline Authors:  Pedro Prudêncio and Leonardo G. Guilgur, date: 05/05/2015, view: 18172, Q&A: 0
The FLP/FRT system is a site-directed recombination technology based on the targeting of a recombination enzyme (flipase - FLP) to specific DNA regions designated as flipase recognition target (FRT) sites. Initially identified in Saccharomyces cerevisiae, the yeast FLP-enzyme and its FRT recombination targets were successfully transferred ...
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Neuroscience

ELISA Detection of Endogenous Serum Albumin in the Mouse Brain: A Measure of Extravasation Following Brain Injury Authors:  Shotaro Michinaga and Yutaka Koyama, date: 05/05/2015, view: 6277, Q&A: 0
After stroke and brain contusion, serum proteins extravasate into nerve tissue through disrupted blood-brain barrier (BBB). Because extravasations of serum proteins result in vasogenic brain edema, serum albumin level in the brain is an indicator of BBB disruption and brain edema after brain insults. In this protocol, extravasation of endogenous ...
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Plant Science

An Assay to Test Manganese Tolerance in Arabidopsis Authors:  Magdalena Migocka and Robert Biskup, date: 05/05/2015, view: 5415, Q&A: 0
Manganese (Mn) is an essential nutrient required for the catalytic or regulatory function of several cellular enzymes. However, excessive Mn concentrations in plant tissues are toxic to plant cells as they negatively affect enzymatic activities, lead to oxidative stress and disturb the uptake and distribution of other essential mineral elements ...
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Quantification of Ralstonia solanacearum Colonization of Potato Germplasm Using Luminescence Authors:  Andrea Paola Zuluaga, Núria S. Coll and Marc Valls, date: 05/05/2015, view: 6562, Q&A: 0
We have developed an unsophisticated, non-disruptive and accurate method for evaluation of pathogen colonization in planta. In this protocol we use a Ralstonia solanacearum (R. solanacearum) UY031 strain genetically modified to constitutively generate light from a synthetic luxCDABE operon stably inserted in its ...
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A Protocol to Measure the Cytoplasmic Calcium in Arabidopsis Guard Cells Authors:  Li Li, Feng Lin, Yana Qu and Qun Zhang, date: 05/05/2015, view: 6634, Q&A: 0
Cytoplasmic calcium ([Ca2+]cyt) acts as a stimulus-induced second messenger in multiple signal transduction cascades (Allen et al., 1999). In plant cells, a dramatic and readily assayed response to stimulus is the change of stomatal aperture. Changes in [Ca2+]cyt of stomatal guard cells were ...
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Arabidopsis Metabolome Analysis Using Infusion ESI FT-ICR/MS Authors:  Reiko Motohashi, Masakazu Satou, Fumiyoshi Myouga, Akira Oikawa and Daisaku Ohta, date: 05/05/2015, view: 5183, Q&A: 0
We made the method for Arabidopsis metabolome analysis based on direct-infusion Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR/MS) (IonSpec). This method was sufficiently applied to metabolic phenotyping of Arabidopsis. This method is simple in that after homogenizing samples, powdered samples are dissolved ...
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Chlorophyll Fluorescence Measurements in Arabidopsis Plants Using a Pulse-amplitude-modulated (PAM) Fluorometer Authors:  Reiko Motohashi and Fumiyoshi Myouga, date: 05/05/2015, view: 11468, Q&A: 0
In this protocol, to analyze PSII activity in photosynthesis, we measure the Fv/Fm (Fv=Fm ± Fo) value (Fo and Fm are the minimum and maximum values of chlorophyll fluorescence of dark-adapted leaves, respectively). Fv/Fm is a reliable marker of photo- inhibition (Krause et al., 1988). Chlorophyll fluorescence in leaves was measured at ...
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MIFE Technique-based Screening for Mesophyll K+ Retention for Crop Breeding for Salinity Tolerance Authors:  Honghong Wu, Lana Shabala, Meixue Zhou and Sergey Shabala, date: 05/05/2015, view: 5880, Q&A: 0
Potassium is known as a rate-limiting factor for crop yield and plays an important role in plants response under abiotic stresses. Recently, cytosolic K+ retention ability in leaf mesophyll has emerged as an important component of plant salt tolerance mechanism (Wu et al., 2013; Wu et al., 2014; Wu et al., ...
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The Use of a Dexamethasone-inducible System to Synchronize Xa21 Expression to Study Rice Immunity Authors:  Daniel F. Caddell, Tong Wei, Chang-Jin Park and Pamela C. Ronald, date: 05/05/2015, view: 8098, Q&A: 0
Inducible gene expression systems offer researchers the opportunity to synchronize target gene expression at particular developmental stages and in particular tissues. The glucocorticoid receptor (GR), a vertebrate steroid receptor, has been well adopted for this purpose in plants. To generate steroid-inducible plants, a construct of GAL4-binding ...
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Stem Cell

Glioma Associated Stem Cells (GASCs) Isolation and Culture Authors:  Evgenia Bourkoula, Damiano Mangoni, Federica Caponnetto, Tamara Ius, Miran Skrap, Antonio Paolo Beltrami and Daniela Cesselli, date: 05/05/2015, view: 5704, Q&A: 0
Glioma Associated Stem Cells (GASCs) represent a population of non-tumorigenic multipotent stem cells hosted in the microenvironment of human gliomas. In vitro, these cells are able, through the release of exosomes, to increase the biological aggressiveness of glioma-initiating cells. The clinical importance of this finding is supported ...
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