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Volume:4 Issue:12
June 20, 2014
Cancer Biology
Lysosomal Stability Assay
Authors:
Nikolaj H. T. Petersen
Nikolaj H. T. Petersen
Affiliation:
Orphazyme ApS, Orphazyme ApS, Copenhagen, Denmark
For correspondence:
nhtp@orphazyme.com
Bio-protocol author page:
a1442
,
Thomas Kirkegaard
Thomas Kirkegaard
Affiliation:
Orphazyme ApS, Orphazyme ApS, Copenhagen, Denmark
Bio-protocol author page:
a1443
and
Marja Jäättelä
Marja Jäättelä
Affiliation:
Cell Death and Metabolism Department, Danish Cancer Society Research Center, Copenhagen, Denmark
Bio-protocol author page:
a1444
,
date: 6/20/2014, 4999 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1162
[Abstract] This assay makes use of the dye Acridine Orange (AO) to determine the stability of lysosomes in living cells upon exposure to a confocal microscope laser.
Figure 1. Acridine Orange
AO is a lipophilic amine that readily diffuses into cells (Figure 1). ...
Article identifier: bio-protocol.org/e1162
Cell Biology
EML Erythroid and Neutrophil Differentiation Protocols
Authors:
Cristina Pina
Cristina Pina
Affiliation:
Department of Haematology, University of Cambridge, Cambridge, UK
For correspondence:
cp533@medschl.cam.ac.uk
Bio-protocol author page:
a1418
,
Cristina Fugazza
Cristina Fugazza
Affiliation:
Dipartimento de Biotecnologie e Bioscienze, Universita degli Studi de Milano-Bicocca, Milan, Italy
Bio-protocol author page:
a1419
and
Tariq Enver
Tariq Enver
Affiliation:
UCL Cancer Institute, University College, London, UK
Bio-protocol author page:
a1420
,
date: 6/20/2014, 3814 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1151
[Abstract] Erythroid-Myeloid-Lymphoid cells (EML) are a multipotent haematopoietic cell line of mouse bone marrow origin capable of long-term maintenance
in vitro
in the presence of SCF (stem cell factor) (Tsai
et al.
, 1994). The self-renewal capacity of the EML ...
Article identifier: bio-protocol.org/e1151
Immunology
Bone Marrow Derived Eosinophil Cultures
Authors:
Thomas X. Lu
Thomas X. Lu
Affiliation:
Division of Pediatrics, Cincinnati Children's Hospital Medical Center / University of Cincinnati, Cincinnati, USA
Bio-protocol author page:
a1440
and
Marc E. Rothenberg
Marc E. Rothenberg
Affiliation:
Division of Pediatrics, Cincinnati Children's Hospital Medical Center / University of Cincinnati, Cincinnati, USA
For correspondence:
marc.rothenberg@cchmc.org
Bio-protocol author page:
a1441
,
date: 6/20/2014, 3730 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1161
[Abstract] Eosinophils are multifunctional effector cells implicated in the pathogenesis of a variety of diseases including asthma, eosinophil gastrointestinal disorders and helminth infection. Mouse bone marrow derived progenitor cells can be differentiated into ...
Article identifier: bio-protocol.org/e1161
Microbiology
A SYBR Green-based Real Time RT-PCR Assay for Detection of the Emerging H7N9 Virus
Authors:
Zheng Zhu
Zheng Zhu
Affiliation:
Institute of Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, China
For correspondence:
zhengzhu@jscdc.cn
Bio-protocol author page:
a1416
and
Lunbiao Cui
Lunbiao Cui
Affiliation:
Institute of Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, China
Bio-protocol author page:
a1417
,
date: 6/20/2014, 2650 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1150
[Abstract] Most recently a novel avian-origin influenza A (H7N9) virus emerged in China and has been associated with lots of human infection and fatal cases. Molecular diagnostic methods are thus urgently needed in public health laboratories. We developed a SYBR ...
Article identifier: bio-protocol.org/e1150
Determination of Oxygen Respiration Rates in Wetted Developmentally Arrested Spores of
Streptomyces
Species
Authors:
Marco Fischer
Marco Fischer
Affiliation:
Department of Microbiology, Martin-Luther University Halle-Wittenberg, Halle (Saale), Germany
Bio-protocol author page:
a1422
,
Dörte Falke
Dörte Falke
Affiliation:
Department of Microbiology, Martin-Luther University Halle-Wittenberg, Halle (Saale), Germany
Bio-protocol author page:
a1423
and
R. Gary Sawers
R. Gary Sawers
Affiliation:
Department of Microbiology, Martin-Luther University Halle-Wittenberg, Halle (Saale), Germany
For correspondence:
gary.sawers@mikrobiologie.uni-halle.de
Bio-protocol author page:
a1424
,
date: 6/20/2014, 2959 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1153
[Abstract]
Streptomyces
species produce spores, which, while not as robust as endospores of
Bacillus
or
Clostridium
species, are capable of surviving for months or even years (Hopwood, 2006). During this time these spores remain viable, surviving by slowly degrading ...
Article identifier: bio-protocol.org/e1153
Rapid Nitrate Reduction Assay with Intact Microbial Cells or Spores
Authors:
Marco Fischer
Marco Fischer
Affiliation:
Department of Microbiology, Martin-Luther University Halle-Wittenberg, Halle (Saale), Germany
Bio-protocol author page:
a1422
,
Dörte Falke
Dörte Falke
Affiliation:
Department of Microbiology, Martin-Luther University Halle-Wittenberg, Halle (Saale), Germany
Bio-protocol author page:
a1423
and
R. Gary Sawers
R. Gary Sawers
Affiliation:
Department of Microbiology, Martin-Luther University Halle-Wittenberg, Halle (Saale), Germany
For correspondence:
gary.sawers@mikrobiologie.uni-halle.de
Bio-protocol author page:
a1424
,
date: 6/20/2014, 2847 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1154
[Abstract] Many microorganisms have the capacity to use nitrate as a respiratory electron acceptor. Reduction of nitrate is catalyzed by a multi-subunit nitrate reductase that is often located associated with the cytoplasmic membrane and has its active site oriented ...
Article identifier: bio-protocol.org/e1154
Analysis of Mycobacterial Protein Secretion
Authors:
Alka Mehra
Alka Mehra
Affiliation:
Department of Medicine, Division of Infectious Diseases, New York University School of Medicine, New York, USA
For correspondence:
alkam08@gmail.com
Bio-protocol author page:
a1435
and
Jennifer A. Philips
Jennifer A. Philips
Affiliation:
Department of Medicine, Division of Infectious Diseases, New York University School of Medicine, New York, USA
For correspondence:
jphilips@dom.wustl.edu
Bio-protocol author page:
a1436
,
date: 6/20/2014, 4317 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1159
[Abstract]
Mycobacterium tuberculosis
(Mtb) is the causative agent of tuberculosis. Analysis of proteins secreted by Mtb has been of interest to the field of tuberculosis research since certain secreted proteins interact with the host to promote virulence, while ...
Article identifier: bio-protocol.org/e1159
Plant Science
Protoplast Preparation and Determination of Cell Death
Author:
Antoine Danon
Antoine Danon
Affiliation:
UMR 8226, Laboratoire de Biologie Moléculaire et Cellulaire des Eucaryotes, Sorbonne Universités, UPMC Univ Paris 06, Centre National de la Recherche Scientifique, Institut de Biologie Physico-Chimique, Paris, France
For correspondence:
antoine.danon@ibpc.fr
Bio-protocol author page:
a1415
,
date: 6/20/2014, 4423 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1149
[Abstract] The protoplasts assay constitutes a powerful tool that allows an easy uptake of active agents and a precise quantification of cell death induction in different populations. Our study showed that the basal level of cell death in our controls is low and ...
Article identifier: bio-protocol.org/e1149
MTV1 Pull-down Assay in
Arabidopsis
Author:
Michael Sauer
Michael Sauer
Affiliation:
Institute of Biochemistry and Biology, Plant Physiology, University of Potsdam, Potsdam, Germany
For correspondence:
michael.sauer@uni-potsdam.de
Bio-protocol author page:
a1421
,
date: 6/20/2014, 3506 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1152
[Abstract] This protocol is an example of how to analyse suspected interactions between proteins using a pull-down assay (Sauer
et al.
, 2013). A bait protein of interest (in this case, MTV1 of
Arabidopsis thaliana
) is fused to a GST tag and expressed in bacteria. ...
Article identifier: bio-protocol.org/e1152
Deflagellation and Regeneration in
Chlamydomonas
Author:
William Dentler
William Dentler
Affiliation:
Department of Molecular Biosciences, University of Kansas, Lawrence, USA
For correspondence:
wdent@ku.edu
Bio-protocol author page:
a551
,
date: 6/20/2014, 4291 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1155
[Abstract] Eukaryotic cilia/flagella are one of the only cellular structures that can be removed without injuring cells, can be highly purified for biochemical analysis, and, in many cells, can be completely reassembled within 90 minutes. Following amputation, ...
Article identifier: bio-protocol.org/e1155
Ciliary and Flagellar Membrane Vesicle (Ectosome) Purification
Author:
William Dentler
William Dentler
Affiliation:
Department of Molecular Biosciences, University of Kansas, Lawrence, USA
For correspondence:
wdent@ku.edu
Bio-protocol author page:
a551
,
date: 6/20/2014, 3369 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1156
[Abstract] Eukaryotic cilia/flagella are ideal organelles for the analysis of membrane trafficking, membrane assembly, and the functions of a variety of signal transduction molecules. Cilia are peninsular organelles and the membrane lipids, membrane proteins, and ...
Article identifier: bio-protocol.org/e1156
In organello
Protein Synthesis
Authors:
Malgorzata Kwasniak-Owczarek
Malgorzata Kwasniak-Owczarek
Affiliation:
Department of Biotechnology, University of Wroclaw, Wroclaw, Poland
Bio-protocol author page:
a1425
and
Hanna Janska
Hanna Janska
Affiliation:
Department of Biotechnology, University of Wroclaw, Wroclaw, Poland
For correspondence:
janska@ibmb.uni.wroc.pl
Bio-protocol author page:
a1426
,
date: 6/20/2014, 3519 views,
1 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1157
[Abstract]
In organello
protein synthesis method allows the analysis of mitochondrial translation products. The principle of this method relies on incubation of isolated intact mitochondria with radiolabeled amino acids such as
35
S methionine. After protein synthesis, ...
Article identifier: bio-protocol.org/e1157
Determination of Luciferase Activity in
Arabidopsis
seedling
Authors:
Mohan TC
Mohan TC
Affiliation:
Department of Plant Molecular Genetics, Centro Nacional de Biotecnología, Madrid, Spain
Bio-protocol author page:
a1437
,
Gabriel Castrillo
Gabriel Castrillo
Affiliation:
Department of Plant Molecular Genetics, Centro Nacional de Biotecnología, Madrid, Spain
Bio-protocol author page:
a1438
and
Antonio Leyva
Antonio Leyva
Affiliation:
Department of Plant Molecular Genetics, Centro Nacional de Biotecnología, Madrid, Spain
For correspondence:
aleyva@cnb.csic.es
Bio-protocol author page:
a1439
,
date: 6/20/2014, 3698 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1160
[Abstract] There are different direct and indirect methods available to study gene expression in plant systems. In this protocol we describe a modified expression assay using transgenic plants expressing the luciferase gene under the control of phosphate transporter ...
Article identifier: bio-protocol.org/e1160
Systems Biology
ChIP-Seq in
Candida albicans
Authors:
Sadri Znaidi
Sadri Znaidi
Affiliation:
Genomes and Genetics Department, Institut Pasteur, Paris, France
For correspondence:
sadri.znaidi@pasteur.fr
Bio-protocol author page:
a1427
,
Caroline Proux
Caroline Proux
Affiliation:
Genomes and Genetics Department, Institut Pasteur, Paris, France
Bio-protocol author page:
a1428
,
Sandra Weber
Sandra Weber
Affiliation:
Biochemistry Department, Institute for Research in Immunology and Cancer and University of Montreal, Montreal, Canada
Bio-protocol author page:
a1429
,
Simon Drouin
Simon Drouin
Affiliation:
Department of Medicine, Clinical Research Institute of Montreal and University of Montreal, Montreal, Canada
Bio-protocol author page:
a1430
,
François Robert
François Robert
Affiliation:
Department of Medicine, Clinical Research Institute of Montreal and University of Montreal, Montreal, Canada
Bio-protocol author page:
a1431
,
Martine Raymond
Martine Raymond
Affiliation:
Biochemistry Department, Institute for Research in Immunology and Cancer and University of Montreal, Montreal, Canada
Bio-protocol author page:
a1432
,
Jean-Yves Coppée
Jean-Yves Coppée
Affiliation:
Genomes and Genetics Department, Institut Pasteur, Paris, France
Bio-protocol author page:
a1433
and
Christophe d’Enfert
Christophe d’Enfert
Affiliation:
Genomes and Genetics Department, Institut Pasteur, Paris, France
For correspondence:
christophe.denfert@pasteur.fr
Bio-protocol author page:
a1434
,
date: 6/20/2014, 5654 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1158
[Abstract] Systems biology approaches can be used to study the regulatory interactions occurring between many components of the biological system at the whole-genome level and decipher the circuitries implicated in the regulation of cellular processes, including ...
Article identifier: bio-protocol.org/e1158
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