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About Us
For Authors
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Preparation Guidelines
Submit Manuscript
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Editorial Criteria
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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
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Biochemistry
FRET-based Stoichiometry Measurements of Protein Complexes
in vitro
Authors:
Francesca Mattiroli
,
Yajie Gu
and
Karolin Luger
,
date:
02/05/2018,
view:
5840,
Q&A:
0
For a complete understanding of biochemical reactions, information on complex stoichiometry is essential. However, measuring stoichiometry is experimentally challenging. Our lab has developed a FRET-based assay to study protein complex stoichiometry
in vitro
. This assay, also known as Job plot, is set up as a continuous variation of the ...
More >>
Cell Biology
Tracking Endocytosis and Intracellular Trafficking of Epitope-tagged Syntaxin 3 by Antibody Feeding in Live, Polarized MDCK Cells
Authors:
Adrian J. Giovannone
,
Elena Reales
,
Pallavi Bhattaram
,
Alberto Fraile-Ramos
and
Thomas Weimbs
,
date:
02/05/2018,
view:
5830,
Q&A:
0
The uptake and trafficking of cell surface receptors can be monitored by a technique called ‘antibody-feeding’ which uses an externally applied antibody to label the receptor on the surface of cultured, live cells. Here, we adapt the traditional antibody-feeding experiment to polarized epithelial cells (Madin-Darby Canine Kidney) grown on ...
More >>
Ex vivo
Analysis of Lipolysis in Human Subcutaneous Adipose Tissue Explants
Authors:
Pauline Decaunes
,
Anne Bouloumié
,
Mikael Ryden
and
Jean Galitzky
,
date:
02/05/2018,
view:
5083,
Q&A:
0
Most studies of human adipose tissue (AT) metabolism and functionality have been performed
in vitro
on isolated mature adipocyte or
in situ
using the microdialysis technique (Lafontan, 2012). However, these approaches have several limitations. The use of mature isolated adipocytes is limiting as adipocytes are not in their ...
More >>
Measurement of Lysosomal Size and Lysosomal Marker Intensities in Adult
Caenorhabditis elegans
Authors:
Julie M. Huynh
,
Hope Dang
and
Hanna Fares
,
date:
02/05/2018,
view:
4617,
Q&A:
0
Assays have been developed to study trafficking in various tissues of
Caenorhabditis elegans
. Adult
C. elegans
intestinal cells are large and have extensive endocytic networks, thus making them a good system for deciphering the endocytic pathway using live imaging techniques. However, the presence of auto-fluorescent gut granules ...
More >>
Analysis of Chromosome Condensation/Decondensation During Mitosis by EdU Incorporation in
Nigella damascena
L. Seedling Roots
Author:
Eugene V. Sheval
,
date:
02/05/2018,
view:
3944,
Q&A:
0
To investigate the chromosome dynamics during mitosis, it is convenient to mark the discrete chromosome foci and then analyze their spatial rearrangements during prophase condensation and telophase decondensation. To label the chromosome regions in plant chromosomes, we incorporated the synthetic nucleotide, 5-ethynyl-2’-deoxyuridine (EdU), which ...
More >>
Developmental Biology
How to Catch a Smurf? – Ageing and Beyond…
In vivo
Assessment of Intestinal Permeability in Multiple Model Organisms
Authors:
Raquel R. Martins
,
Andrew W. McCracken
,
Mirre J. P. Simons
,
Catarina M. Henriques
and
Michael Rera
,
date:
02/05/2018,
view:
7574,
Q&A:
1
The Smurf Assay (SA) was initially developed in the model organism
Drosophila melanogaster
where a dramatic increase of intestinal permeability has been shown to occur during aging (Rera
et al.
, 2011). We have since validated the protocol in multiple other model organisms (Dambroise
et al.
, 2016) and have utilized the ...
More >>
Preparation of Precisely Oriented Cryosections of Undistorted
Drosophila
Wing Imaginal Discs for High Resolution Confocal Imaging
Authors:
Samuel Petshow
and
Marcel Wehrli
,
date:
02/05/2018,
view:
6041,
Q&A:
0
The combination of immunofluorescence and laser scanning confocal microscopy (LSM) is essential to high-resolution detection of molecular distribution in biological specimens. A frequent limitation is the need to image deep inside a tissue or in a specific plane, which may be inaccessible due to tissue size or shape. Recreating high-resolution 3D ...
More >>
Microbiology
Extraction of DNA from Murine Fecal Pellets for Downstream Phylogenetic Microbiota Analysis by Next-generation Sequencing
Authors:
Elien Eeckhout
and
Andy Wullaert
,
date:
02/05/2018,
view:
8163,
Q&A:
2
Mouse models are widely used to evaluate the potential impact of the gut microbial composition on health and disease. Standardized protocols for sampling and storing murine feces, as well as for extracting DNA from these fecal pellets are needed to limit experimental variation between different studies. Both efficient lysis of the microbiota and ...
More >>
Quantification of
Neisseria meningitidis
Adherence to Human Epithelial Cells by Colony Counting
Authors:
Sara Sigurlásdóttir
,
Sunil D Saroj
,
Olaspers Sara Eriksson
,
Jens Eriksson
and
Ann-Beth Jonsson
,
date:
02/05/2018,
view:
5260,
Q&A:
0
To cause an infection, the human specific pathogen
Neisseria meningitides
must first colonize the nasopharynx. Upon tight interaction with the mucosal epithelium,
N. meningitidis
may cross the epithelial cellular barrier, reach the bloodstream and cause sepsis and/or meningitis. Since
N. meningitidis
niche is restricted ...
More >>
Immunofluorescence Analysis of Human Endocervical Tissue Explants Infected with
Neisseria gonorrhoeae
Authors:
Liang-Chun Wang
,
Qian Yu
,
Daniel C. Stein
and
Wenxia Song
,
date:
02/05/2018,
view:
4307,
Q&A:
0
Colonization and penetration of the epithelium is the infection-initiating route of mucosal pathogens. The epithelium counteracts infection by eliciting host cell responses while maintaining the mucosal barrier function. The obligate human sexually transmitted bacterium
Neisseria gonorrhoeae
, or
gonococcus
(GC) infects the female ...
More >>
Molecular Biology
Determination of DNA Damage in the Retina Photoreceptors of
Drosophila
Authors:
Damulewicz Milena
and
Pyza Elzbieta
,
date:
02/05/2018,
view:
4316,
Q&A:
0
The retina is sensitive for light damages, because of direct light exposure, especially intense blue and UV light, which increase level of ROS and other toxic phototransduction products in photoreceptor cells. In our previous work (Damulewicz
et al.
, 2017a and 2017b), we used 8-oxo-deoxyguanosine (8-OHdG) as a marker for oxidative stress ...
More >>
Measuring Nucleosome Assembly Activity
in vitro
with the Nucleosome Assembly and Quantification (NAQ) Assay
Authors:
Francesca Mattiroli
,
Yajie Gu
and
Karolin Luger
,
date:
02/05/2018,
view:
4501,
Q&A:
0
Nucleosomes organize the eukaryotic genome into chromatin. In cells, nucleosome assembly relies on the activity of histone chaperones, proteins with high binding affinity to histones. At least a subset of histone chaperones promotes histone deposition
in vivo
. However, it has been challenging to characterize this activity, due to the lack ...
More >>
Medaka-microinjection with an Upright Microscope
Authors:
Yu Murakami
and
Masato Kinoshita
,
date:
02/05/2018,
view:
4977,
Q&A:
0
We described a simple method for microinjecting DNA/RNA/Protein solutions into medaka eggs under an upright microscope. Medaka is an excellent vertebrate model for reverse genetics, because of its daily spawning, short generation time, and egg transparency. These features enable us to efficiently perform functional genomic analyses of transgenic ...
More >>
Immunoprecipitation of Tri-methylated Capped RNA
Authors:
Karen E. Hayes
,
Jamie A. Barr
,
Mingyi Xie
,
Joan A. Steitz
and
Ivan Martinez
,
date:
02/05/2018,
view:
6044,
Q&A:
0
Cellular quiescence (also known as G
0
arrest) is characterized by reduced DNA replication, increased autophagy, and increased expression of cyclin-dependent kinase p27
Kip1
. Quiescence is essential for wound healing, organ regeneration, and preventing neoplasia. Previous findings indicate that microRNAs (miRNAs) play an ...
More >>
Precision Tagging: A Novel Seamless Protein Tagging by Combinational Use of Type II and Type IIS Restriction Endonucleases
Authors:
Zhen Xu
,
Yan-Ning Rui
,
John P. Hagan
and
Dong H. Kim
,
date:
02/05/2018,
view:
5784,
Q&A:
0
Protein tagging is a powerful tool for performing comprehensive analyses of the biological functions of a protein of interest owing to the existence of a wide variety of tags. It becomes indispensable in some cases, such as in tracking protein dynamics in a live cell or adding a peptide epitope due to the lack of optimal antibodies. However, ...
More >>
Neuroscience
Culture and Nucleofection of Postnatal Day 7 Cortical and Cerebellar Mouse Astroglial Cells
Authors:
Malek Chouchane
and
Marcos Romualdo Costa
,
date:
02/05/2018,
view:
4790,
Q&A:
0
Lineage reprogramming of astroglial cells isolated from different brain regions leads to the generation of different neuronal subtypes. This protocol describes the isolation and culture of neocortical and cerebellar astrocytes from postnatal mice. We also present a comprehensive description of the main steps towards successful gene delivery in ...
More >>
Plant Science
Identification of Insertion Site by RESDA-PCR in
Chlamydomonas
Mutants Generated by
AphVIII
Random Insertional Mutagenesis
Authors:
Fantao Kong
and
Yonghua Li-Beisson
,
date:
02/05/2018,
view:
4891,
Q&A:
0
Chlamydomonas reinhardtii
is frequently used as a model organism to study fundamental processes in photosynthesis, metabolism, and flagellar biology. Versatile tool boxes have been developed for this alga (Fuhrmann
et al.
, 1999; Schroda
et al.
, 2000; Schroda, 2006). Among them, forward genetic approach has been ...
More >>
Radioactive Tracer Feeding Experiments and Product Analysis to Determine the Biosynthetic Capability of Comfrey (
Symphytum officinale
) Leaves for Pyrrolizidine Alkaloids
Authors:
Thomas Stegemann
,
Lars H. Kruse
and
Dietrich Ober
,
date:
02/05/2018,
view:
4202,
Q&A:
0
This protocol delivers a method to determine the biosynthetic capability of comfrey leaves for pyrrolizidine alkaloids independently from other organs like roots or flowers.
The protocol applies and combines radioactive tracer experiments with standard and modern techniques like thin layer chromatography (TLC), solid-phase extraction ...
More >>
Investigating Localization of Chimeric Transporter Proteins within Chloroplasts of
Arabidopsis thaliana
Authors:
Susumu Uehara
,
Yasuko Ito-Inaba
and
Takehito Inaba
,
date:
02/05/2018,
view:
6071,
Q&A:
0
In this protocol, we describe a method to design chimeric proteins for specific targeting to the inner envelope membrane (IEM) of
Arabidopsis
chloroplasts and the confirmation of their localization by biochemical analysis. Specific targeting to the chloroplast IEM can be achieved by fusing the protein of interest with a transit peptide ...
More >>
Systems Biology
Extraction and Analysis of Pan-metabolome Polar Metabolites by Ultra Performance Liquid Chromatography–Tandem Mass Spectrometry (UPLC-MS/MS)
Authors:
Dania M. Malik
,
Seth Rhoades
and
Aalim Weljie
,
date:
02/05/2018,
view:
5435,
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 advantageous, as it does not require derivatization procedures and a large diversity in physiochemical characteristics of analytes can be accommodated through a ...
More >>
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