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Volume:3  Issue:22

November 20, 2013

Cell Biology

Harvest and Culture of Mouse Peritoneal Macrophages

Authors: Mingfang Lu
Mingfang LuAffiliation: Internal Medicine, University of Texas Southwestern Medical School, Dallas, TX, USA
Bio-protocol author page: a990
 and Alan W. Varley
Alan W. VarleyAffiliation: Internal Medicine, University of Texas Southwestern Medical School, Dallas, TX, USA
For correspondence: alanwaynevarley@gmail.com
Bio-protocol author page: a991
date: 11/20/2013, 13135 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.976 

[Abstract] Peritoneal macrophages are used as primary macrophages in lots of studies, mainly because they are easy to obtain. Injection of thioglycollate broth i.p. induces inflammatory responses and elicits large numbers of macrophages. This protocol can be used ...

Article identifier: bio-protocol.org/e976

Immunology

Gastric Aspiration Models

Authors: Bruce A. Davidson
Bruce A. DavidsonAffiliation: Department of Anesthesiology, and Pathology and Anatomical Sciences, State University of New York at Buffalo, Buffalo, USA
For correspondence: bdavidso@buffalo.edu
Bio-protocol author page: a1019
 and Ravi Alluri
Ravi AlluriAffiliation: Department of Anesthesiology, and Pathology and Anatomical Sciences, State University of New York at Buffalo, Buffalo, USA
Bio-protocol author page: a1020
date: 11/20/2013, 7446 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.968 

[Abstract] The procedures described below are for producing gastric aspiration pneumonitis in mice with alterations for rats and rabbits described parenthetically. We use 4 different injury vehicles delivered intratracheally to investigate the inflammatory responses ...

Article identifier: bio-protocol.org/e968

Immunocytochemical Detection of Recombinant Biomphalysin on Schistosoma mansoni Sporocysts

Authors: David Duval
David DuvalAffiliation: Ecologie et Evolution des Interactions (2EI), UPVD, CNRS, UMR 5244, Perpignan, France
For correspondence: david.duval@univ-perp.fr
Bio-protocol author page: a1014
Richard Galinier
Richard GalinierAffiliation: Ecologie et Evolution des Interactions (2EI), UPVD, CNRS, UMR 5244, Perpignan, France
Bio-protocol author page: a1015
Julien Portela
Julien PortelaAffiliation: Ecologie et Evolution des Interactions (2EI), UPVD, CNRS, UMR 5244, Perpignan, France
Bio-protocol author page: a1016
Guillaume Mitta
Guillaume Mitta Affiliation: Ecologie et Evolution des Interactions (2EI), UPVD, CNRS, UMR 5244, Perpignan, France
Bio-protocol author page: a1017
 and Benjamin Gourbal
Benjamin GourbalAffiliation: Ecologie et Evolution des Interactions (2EI), UPVD, CNRS, UMR 5244, Perpignan, France
Bio-protocol author page: a1018
date: 11/20/2013, 2903 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.969 

[Abstract] Schistosomiasis, or bilharzia, is a tropical disease caused by worms of the genus Schistosoma which infect about 200 million people. The life cycle of the parasite requires Biomphalaria, a specific genus of freshwater snails, as intermediate. Using an ...

Article identifier: bio-protocol.org/e969

Assessment of Human Dendritic Cell Antigen Uptake by Flow Cytometry

Authors: Ana Luque
Ana LuqueAffiliation: Human Molecular Genetics Department, IDIBELL, L'Hospitalet de Llobregat, Spain
Bio-protocol author page: a995
Sonia Cárdenas-Brito
Sonia Cárdenas-BritoAffiliation: Human Molecular Genetics Department, IDIBELL, L'Hospitalet de Llobregat, Spain
Bio-protocol author page: a996
Rut Olivar
Rut OlivarAffiliation: Human Molecular Genetics Department, IDIBELL, L'Hospitalet de Llobregat, Spain
Bio-protocol author page: a997
 and Josep M. Aran
Josep M. AranAffiliation: Human Molecular Genetics Department, IDIBELL, L'Hospitalet de Llobregat, Spain
For correspondence: jaran@idibell.cat
Bio-protocol author page: a998
date: 11/20/2013, 4766 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.974 

[Abstract] Antigen uptake by dendritic cells is the first key step towards induction of antigen-specific T-cell responses. This flow cytometry-based protocol describes the analysis of dendritic cell uptake of soluble antigens through two different mechanisms: non-specific ...

Article identifier: bio-protocol.org/e974

Microbiology

Separation and Detection of Phosphorylated and Nonphosphorylated BvgA, a Bordetella pertussis Response Regulator, in vivo and in vitro

Authors: Qing Chen
Qing ChenAffiliation: Center For Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD, USA
Bio-protocol author page: a1010
Alice Boulanger
Alice BoulangerAffiliation: NIDDK, National Institutes of Health, Bethesda, MD, USA
Bio-protocol author page: a1011
Deborah M. Hinton
Deborah M. HintonAffiliation: NIDDK, National Institutes of Health, Bethesda, MD, USA
For correspondence: dhinton@helix.nih.gov
Bio-protocol author page: a1012
 and Scott Stibitz
Scott StibitzAffiliation: Center For Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD, USA
Bio-protocol author page: a1013
date: 11/20/2013, 4224 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.970 

[Abstract] Protein phosphorylation plays a central role in signal transduction in bacteria. However, separation and detection of the phosphorylated protein from its nonphosphorylated form remain challenging. Here we describe a method to detect phosphorylation of ...

Article identifier: bio-protocol.org/e970

Transport Assays in Aspergillus nidulans

Authors: Emilia Krypotou
Emilia Krypotou Affiliation: Faculty of Biology, University of Athens, Athens, Greece
Bio-protocol author page: a1008
 and George Diallinas
George DiallinasAffiliation: Faculty of Biology, University of Athens, Athens, Greece
For correspondence: diallina@biol.uoa.gr
Bio-protocol author page: a1009
date: 11/20/2013, 2708 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.971 

[Abstract] Transport assays allow the direct kinetic analysis of a specific transporter by measuring apparent Km and Vmax values, and permit the characterization of substrate specificity profiles through competition assays. In this protocol, we describe a rapid ...

Article identifier: bio-protocol.org/e971

Shigella IpaD and IpaB Surface Localizations

Authors: Lionel Schiavolin
Lionel SchiavolinAffiliation: Molecular Bacteriology Lab, Université Libre de Bruxelles, Anderlecht, Belgium
For correspondence: lschiavo@ulb.ac.be
Bio-protocol author page: a992
Alaeddine Meghraoui
Alaeddine MeghraouiAffiliation: Molecular Bacteriology Lab, Université Libre de Bruxelles, Anderlecht, Belgium
Bio-protocol author page: a993
 and Abdelmounnaim Allaoui
Abdelmounnaim AllaouiAffiliation: Molecular Bacteriology Lab, Université Libre de Bruxelles, Anderlecht, Belgium
Bio-protocol author page: a994
date: 11/20/2013, 3459 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.975 

[Abstract] Shigella uses a type III secretion system to invade host cell and to cause disease. Secretion control and insertion of a translocation pore into cell membrane are critical steps for pathogenesis and are tightly linked to the formation of the needle tip ...

Article identifier: bio-protocol.org/e975

Assay to Evaluate Vascular Permeability Induction in Mice

Authors: Henry Puerta-Guardo
Henry Puerta-GuardoAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a980
Arturo Raya-Sandino
Arturo Raya-SandinoAffiliation: Department of Physiolology, Biophysics and Neurosciences, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a981
Lorenza González-Mariscal
Lorenza González-MariscalAffiliation: Department of Physiolology, Biophysics and Neurosciences, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a982
Victor H. Rosales
Victor H. RosalesAffiliation: General Services Laboratory, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a983
José Ayala-Dávila
José Ayala-DávilaAffiliation: Department of Physiolology, Biophysics and Neurosciences, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a984
Bibiana Chávez-Mungía
Bibiana Chávez-MungíaAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a985
Daniel Martínez-Fong
Daniel Martínez-FongAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a986
Fernando Medina
Fernando MedinaAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a987
Juan E. Ludert
Juan E. LudertAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
For correspondence: jludert@cinvestav.mx
Bio-protocol author page: a988
 and Rosa María del Angel
Rosa María del AngelAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
For correspondence: rmangel@cinvestav.mx
Bio-protocol author page: a989
date: 11/20/2013, 9127 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.977 

[Abstract] Dengue virus infection usually courses as a benign self-limited fever, called dengue fever. However, on occasions it can progress to a life-threatening complication known as severe dengue (SD). A hallmark of SD is a sharp increase in vascular permeability. ...

Article identifier: bio-protocol.org/e977

Neuroscience

Cell Cycle Analysis in the Vertebrate Brain Using Immunolabeled Fresh Cell Nuclei

Authors: Noelia Lopez-Sanchez
Noelia Lopez-SanchezAffiliation: Molecular, Cellular and Developmental Neurobiology, Cajal Institute, CSIC, Madrid, Spain
Bio-protocol author page: a999
 and Jose M. Frade
Jose M. FradeAffiliation: Molecular, Cellular and Developmental Neurobiology, Cajal Institute, CSIC, Madrid, Spain
For correspondence: frade@cajal.csic.es
Bio-protocol author page: a1000
date: 11/20/2013, 7123 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.973 

[Abstract] Flow cytometry, a standard technique used for quantitative analysis of isolated cells, is routinely employed by immunologists and oncologists to study DNA content, protein expression, and other functional parameters in blood and tumor cells. Unfortunately, ...

Article identifier: bio-protocol.org/e973

Plant Science

Mapping and Analysis of Illumina Reads for Transcriptome of Medicago Truncatula During the Early Organogenesis of the Nodule

Authors: Alexandre Boscari
Alexandre BoscariAffiliation: Institut National de la Recherche Agronomique, Unité Mixte de Recherche 1355, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7254, and Université de Nice-Sophia Antipolis, Unité Mixte de Recherche Institut Sophia Agrobiotech, Sophia Antipolis, France
For correspondence: alexandre.boscari@sophia.inra.fr
Bio-protocol author page: a504
Alberto Ferrarini
Alberto FerrariniAffiliation: Università degli Studi di Verona, Dipartimento di Biotecnologie, Centro di Genomica Funzionale, Verona, Italy
Bio-protocol author page: a1025
Jennifer del Giudice
Jennifer del GiudiceAffiliation: Institut National de la Recherche Agronomique, Unité Mixte de Recherche 1355, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7254, and Université de Nice-Sophia Antipolis, Unité Mixte de Recherche Institut Sophia Agrobiotech, Sophia Antipolis, France
Bio-protocol author page: a1026
Luca Venturini
Luca VenturiniAffiliation: Università degli Studi di Verona, Dipartimento di Biotecnologie, Centro di Genomica Funzionale, Verona, Italy
Bio-protocol author page: a1027
Massimo Delledone
Massimo DelledoneAffiliation: Università degli Studi di Verona, Dipartimento di Biotecnologie, Centro di Genomica Funzionale, Verona, Italy
Bio-protocol author page: a1028
 and Alain Puppo
Alain PuppoAffiliation: Institut National de la Recherche Agronomique, Unité Mixte de Recherche 1355, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7254, and Université de Nice-Sophia Antipolis, Unité Mixte de Recherche Institut Sophia Agrobiotech, Sophia Antipolis, France
For correspondence: puppo@unice.fr
Bio-protocol author page: a1029
date: 11/20/2013, 4214 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.966 

[Abstract] Medicago truncatula serves as a model plant for legume genetics and genomics. We used RNA-Seq to characterize the transcriptome during the early organogenesis of the nodule and during its functioning. We generated approximately 135.5 million high-quality ...

Article identifier: bio-protocol.org/e966

Analysis of RNA-protein Interactions Using Electrophoretic Mobility Shift Assay (Gel Shift Assay)

Authors: Saiprasad Goud Palusa
Saiprasad Goud PalusaAffiliation: Department of Biology, Colorado State University, Fort Collins, USA
Bio-protocol author page: a1022
 and Anireddy S. N. Reddy
Anireddy S. N. ReddyAffiliation: Department of Biology, Colorado State University, Fort Collins, USA
For correspondence: reddy@colostate.edu
Bio-protocol author page: a1023
date: 11/20/2013, 10892 views, 4 Q&A
DOI: https://doi.org/10.21769/BioProtoc.967 

[Abstract] RNA binding proteins (RBPs) play a crucial role in regulating gene expression at the post-transcriptional level at multiple steps including pre-mRNA splicing, polyadenylation, mRNA stability, mRNA localization and translation. RBPs regulate these processes ...

Article identifier: bio-protocol.org/e967

Cotton Ovules Culture and Analysis

Authors: Jiafu Tan
Jiafu TanAffiliation: National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, China
Bio-protocol author page: a1001
Fenglin Deng
Fenglin DengAffiliation: National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, China
Bio-protocol author page: a1002
Wenxin Tang
Wenxin TangAffiliation: National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, China
Bio-protocol author page: a1003
Jie Han
Jie HanAffiliation: National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, China
Bio-protocol author page: a1004
Guo Kai
Guo KaiAffiliation: National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, China
Bio-protocol author page: a1005
Lili Tu
Lili TuAffiliation: National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, China
Bio-protocol author page: a1006
 and Xianlong Zhang
Xianlong ZhangAffiliation: National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, China
For correspondence: xlzhang@mail.hzau.edu.cn
Bio-protocol author page: a1007
date: 11/20/2013, 2873 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.972 

[Abstract] The cotton ovules culture was innovated by Beasley and Ting (1973), and named after them. It is a convenient system to analyze the effect of chemical or environmental treatment on fiber development directly on ovules. This protocol was generated according ...

Article identifier: bio-protocol.org/e972

Shikimate Hydroxycinnamoyl Transferase (HCT) Activity Assays in Populus nigra

Authors: Igor Cesarino
Igor CesarinoAffiliation: Department of Plant Systems Biology / Department of Plant Biotechnology and Bioinformatics, VIB / Ghent University, Gent, Belgium
Bio-protocol author page: a975
Ruben Vanholme
Ruben VanholmeAffiliation: Department of Plant Systems Biology / Department of Plant Biotechnology and Bioinformatics, VIB / Ghent University, Gent, Belgium
Bio-protocol author page: a976
Geert Goeminne
Geert GoeminneAffiliation: Department of Plant Systems Biology / Department of Plant Biotechnology and Bioinformatics, VIB / Ghent University, Gent, Belgium
Bio-protocol author page: a977
Bartel Vanholme
Bartel VanholmeAffiliation: Department of Plant Systems Biology / Department of Plant Biotechnology and Bioinformatics, VIB / Ghent University, Gent, Belgium
Bio-protocol author page: a978
 and Wout Boerjan
Wout BoerjanAffiliation: Department of Plant Systems Biology / Department of Plant Biotechnology and Bioinformatics, VIB / Ghent University, Gent, Belgium
For correspondence: wout.boerjan@psb.vib-ugent.be
Bio-protocol author page: a979
date: 11/20/2013, 4557 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.978 

[Abstract] Lignin is a complex phenolic polymer deposited in secondarily-thickened plant cell walls. The polymer is mainly derived from the three primary monolignols: p-coumaryl, coniferyl and sinapyl alcohol which give rise to p-hydroxyphenyl, guaiacyl and syringyl ...

Article identifier: bio-protocol.org/e978

Bimolecular Fluorescence Complementation (BIFC) Protocol for Rice Protoplast Transformation

Authors: Kun Wang
Kun WangAffiliation 1: State Key Laboratory of Hybrid Rice, College of Life Sciences, Wuhan University, Wuhan, China
Affiliation 2: Wuhan Botanical Garden, Chinese Academy of Science, Wuhan, China
Bio-protocol author page: a973
Ying Liu
Ying LiuAffiliation: State Key Laboratory of Hybrid Rice, College of Life Sciences, Wuhan University, Wuhan, China
Bio-protocol author page: a1825
 and Shaoqing Li
Shaoqing LiAffiliation: State Key Laboratory of Hybrid Rice, College of Life Sciences, Wuhan University, Wuhan, China
For correspondence: shaoqingli@whu.edu.cn
Bio-protocol author page: a974
date: 11/20/2013, 4067 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.979 

[Abstract] After the plant cells are removed the cell walls by digestive enzyme, the plant protoplasts still have good cell activity. The protoplasts can be used to transiently express proteins of target genes in living plant cells through polyethylene glycol (PEG) ...

Article identifier: bio-protocol.org/e979

Binding Assay of Cytosolic Proteins to the Cytoskeleton

Authors: Stefano Del Duca
Stefano Del DucaAffiliation: Dipartimento di Scienze Biologiche, Geologiche ed Ambientali, University of Bologna, Bologna, Italy
For correspondence: stefano.delduca@unibo.it
Bio-protocol author page: a971
 and Giampiero Cai
Giampiero CaiAffiliation: Dipartimento Scienze della Vita, University of Siena, Siena, Italy
Bio-protocol author page: a972
date: 11/20/2013, 3608 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.980 

[Abstract] Many cellular proteins interact with the cytoskeleton (both actin filaments and microtubules), either dynamically or permanently. This interaction is required during different aspects of the cell life, for example during the process of cell division. ...

Article identifier: bio-protocol.org/e980