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About Us
For Authors
Submission Procedure
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
Preparation, FPLC Purification and LC-FT-ICR-MS of Proteins
Authors:
Tyler B. Johnson
,
Jiri Adamec
and
Paul Blum
,
date:
04/05/2020,
view:
1539,
Q&A:
0
High magnetic field Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers provide extremely high mass resolution (resolving power of ~200,000 at 400 m/z) protein detection across a broad mass range, enabling analysis of fine structure of isotopic peak clusters that is missed in other types of mass spectrometers. The protocol ...
More >>
Determination of Flavin Potential in Proteins by Xanthine/Xanthine Oxidase Method
Authors:
Elena Maklashina
and
Gary Cecchini
,
date:
04/05/2020,
view:
1467,
Q&A:
0
This protocol describes a simple xanthine/xanthine oxidase enzymatic equilibration method for determination of the redox potential of a flavin. As an example of the use of this method, we determine the reduction potential of the covalently bound FAD cofactor (
E
m
= -55 mV) in the SdhA flavoprotein subunit of succinate ...
More >>
Measurement of Protein-Protein Interactions through Microscale Thermophoresis (MST)
Authors:
Magnez Romain
,
Bryan Thiroux
,
Morgane Tardy
,
Bruno Quesnel
and
Xavier Thuru
,
date:
04/05/2020,
view:
1768,
Q&A:
0
The binding interactions of PD-1 and PD-L1 have been studied by surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC) over the past few years, but these investigations resulted in controversy regarding the values of the dissociation constant (K
d
) (Freeman
et al.
, 2000). MST is a powerful new method for the ...
More >>
Spectrophotometric Assessment of Heme Oxygenase-1 Activity in
Leishmania
-infected Macrophages
Authors:
Moumita Basu
,
Shriya Saha
and
Anindita Ukil
,
date:
04/05/2020,
view:
1410,
Q&A:
0
Heme oxygenase-1 (HO-1) is a stress responsive enzyme that metabolizes heme and releases free iron, carbon monoxide (CO), and biliverdin (BV), which rapidly undergoes conversion to bilirubin (BL). Estimation of bilirubin is the basis of HO-1 assay. HO-1 activity is widely employed to determine antioxidant response of cells under different ...
More >>
Quantitative Determination of Ca
2+
-binding to Ca
2+
-sensor Proteins by Isothermal Titration Calorimetry
Authors:
Seher Abbas
and
Karl-Wilhelm Koch
,
date:
04/05/2020,
view:
1582,
Q&A:
0
Diverse and complex molecular recognitions are central elements of signal transduction cascades. The strength and nature of these interaction modes can be determined by different experimental approaches. Among those, Isothermal titration calorimetry (ITC) offers certain advantages by providing binding constants and thermodynamic parameters from ...
More >>
Cancer Biology
In vivo
Tumor Growth and Spontaneous Metastasis Assays Using A549 Lung Cancer Cells
Authors:
Lei Qi
,
Teresa Knifley
,
Dava W. Piecoro
,
Piotr Rychahou
,
Jianrong Wu
,
Kathleen L. O’Connor
and
Min Chen
,
date:
04/05/2020,
view:
1935,
Q&A:
0
Metastasis accounts for the majority of cancer related deaths. The genetically engineered mouse (GEM) models and cell line-based subcutaneous and orthotopic mouse xenografts have been developed to study the metastatic process. By using lung cancer cell line A549 as an example, we present a modified protocol to establish the cell line-based ...
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Cell Biology
Flow Cytometry Measurement of Glucocerebrosidase Activity in Human Monocytes
Authors:
Laura P. Hughes
,
Glenda M. Halliday
and
Nicolas Dzamko
,
date:
04/05/2020,
view:
1768,
Q&A:
0
Glucocerebrosidase (GCase) is an important enzyme for the metabolism of glycolipids. GCase enzyme deficiency is implicated in human disease and the efficient measurement of GCase activity is important for evaluating the efficacy of therapeutics targeting this enzyme. Existing approaches to measure GCase activity include whole blood mass ...
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Isolation, Purification and Characterization of Exosomes from Fibroblast Cultures of Skeletal Muscle
Authors:
Diantha van de Vlekkert
,
Xiaohui Qiu
,
Ida Annunziata
and
Alessandra d'Azzo
,
date:
04/05/2020,
view:
1736,
Q&A:
0
Exosomes are dynamic nanovesicles secreted by virtually all cells and are present in all biological fluids. Given their highly heterogeneous content exosomes have been implicated in many physiological and pathological processes that they exert by influencing cell-cell and cell-ECM communication. In recent years an increasing number of methods have ...
More >>
Fractionation of Ovarian Follicles and
in vitro
Oocyte Maturation and Ovulation Assay of
Ciona intestinalis
Type A
Authors:
Shin Matsubara
,
Akira Shiraishi
,
Tomohiro Osugi
,
Tsuyoshi Kawada
and
Honoo Satake
,
date:
04/05/2020,
view:
1481,
Q&A:
0
Ascidians are the closest living relatives of vertebrates (Delsuc
et al.
, 2006; Satoh
et al.
, 2014) and are important for the evolutionary study of the ovarian follicle development including oocyte maturation and ovulation. However, neither the endogenous factors nor the molecular mechanisms underlying the oocyte maturation and ...
More >>
Developmental Biology
Bimolecular Fluorescence Complementation (BiFC) for Studying Sarcomeric Protein Interactions in
Drosophila
Authors:
Océane Marescal
,
Frieder Schöck
and
Nicanor González-Morales
,
date:
04/05/2020,
view:
1759,
Q&A:
0
Protein-protein interactions in
Drosophila
myofibrils are essential for their function and formation. Bimolecular Fluorescence Complementation (BiFC) is an effective method for studying protein interactions and localization. BiFC relies on the reconstitution of a monomeric fluorescent protein from two half-fragments when in proximity. Two ...
More >>
Microbiology
Conjugation Protocol Optimised for
Roseburia inulinivorans
and
Eubacterium rectale
Authors:
Paul O. Sheridan
,
Jennifer C. Martin
and
Karen P. Scott
,
date:
04/05/2020,
view:
1357,
Q&A:
0
Roseburia
and
Eubacterium
species of the human gut microbiota play an important role in the maintaince of human health, partly by producing butyrate, the main energy source of our colonic epithelial cells. However, our knowledge of the biochemistry and physiology of these bacteria has been limited by a lack of genetic ...
More >>
Molecular Biology
Preparation of a Bacteriophage T4-based Prokaryotic-eukaryotic Hybrid Viral Vector for Delivery of Large Cargos of Genes and Proteins into Human Cells
Authors:
Jingen Zhu
,
Pan Tao
,
Marthandan Mahalingam
and
Venigalla B. Rao
,
date:
04/05/2020,
view:
1185,
Q&A:
0
A viral vector that can safely and efficiently deliver large and diverse molecular cargos into cells is the holy grail of curing many human diseases. Adeno-associated virus (AAV) has been extensively used but has a very small capacity. The prokaryotic virus T4 has a large capacity but lacks natural mechanisms to enter mammalian cells. Here, we ...
More >>
Measuring cAMP Specific Phosphodiesterase Activity: A Two-step Radioassay
Authors:
Connor M. Blair
,
Jiayue Ling
and
George S. Baillie
,
date:
04/05/2020,
view:
994,
Q&A:
0
Cyclic nucleotide degrading phosphodiesterase (PDE) enzymes are crucial to the fine tuning of cAMP signaling responses, playing a pivotal role in regulating the temporal and spatial characteristics of discrete cAMP nanodomains and hence the activity of cAMP-effector proteins. As a consequence of orchestrating cAMP homeostasis, dysfunctional PDE ...
More >>
Real-time Fluorescence Measurement of Enterovirus Uncoating
Authors:
Visa Ruokolainen
,
Mira Laajala
and
Varpu Marjomäki
,
date:
04/05/2020,
view:
1217,
Q&A:
0
Viruses need to open,
i.e.
, uncoat, in order to release their genomes for efficient replication and translation. Especially for non-enveloped viruses, such as enteroviruses, the cues leading to uncoating are less well known. The status of the virus has previously been observed mainly by transmission electron microscopy using negative ...
More >>
Neuroscience
A Simple and Fast Battery Test for Phenotypic Characterization of Mice
Authors:
Mariana S. A. Garcia-Gomes
,
Dennis A. Zanatto
,
Pedro K. Yamamoto
,
Danilo Wadt
,
Ana T. F. B. Antiorio
,
Jilma Aleman-Laporte
,
Sandra R. Alexandre-Ribeiro
,
Guilherme A. Marson
,
Cezar Guizzo
,
Silvia M. G. Massironi
,
Maria M. Bernardi
and
Claudia M. C. Mori
,
date:
04/05/2020,
view:
1350,
Q&A:
0
Despite the great number of test batteries already known to assess the behavior of genetically modified and inbred strains of mice, only a few of them focus on basic neurological parameters. The purpose of the battery test proposed is to settle a specific methodology to characterize the phenotype of neurological disease models in mutant or ...
More >>
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