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Volume:4 Issue:22
November 20, 2014
Cancer Biology
Induction of Colitis and Colitis-associated Colorectal Cancer (CAC)
Authors:
Jalaj Gupta
Jalaj Gupta
Affiliation:
Institute for Research in Biomedicine (IRB Barcelona), Barcelona, Spain
For correspondence:
Gupta@gsh.uni-frankfurt.de
Bio-protocol author page:
a1790
and
Angel R. Nebreda
Angel R. Nebreda
Affiliation 1:
Institute for Research in Biomedicine (IRB Barcelona), Barcelona, Spain
Affiliation 2:
Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain
For correspondence:
angel.nebreda@irbbarcelona.org
Bio-protocol author page:
a1791
,
date: 11/20/2014, 4975 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1290
[Abstract] Inflammatory bowel disease (IBD) including Crohn’s disease and ulcerative colitis are characterized by chronic, progressive and relapsing inflammatory disorders. Existing evidence indicate that IBD is associated with a higher risk of developing CAC, ...
Article identifier: bio-protocol.org/e1290
Purification of Tumor-Associated Macrophages (TAM) and Tumor-Associated Dendritic Cells (TADC)
Authors:
Damya Laoui
Damya Laoui
Affiliation 1:
Myeloid Cell Immunology Laboratory, Brussels, Belgium
Affiliation 2:
Lab of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium
Bio-protocol author page:
a1803
,
Eva Van Overmeire
Eva Van Overmeire
Affiliation 1:
Myeloid Cell Immunology Laboratory, Brussels, Belgium
Affiliation 2:
Lab of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium
Bio-protocol author page:
a1804
,
Jiri Keirsse
Jiri Keirsse
Affiliation 1:
Myeloid Cell Immunology Laboratory, Brussels, Belgium
Affiliation 2:
Lab of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium
Bio-protocol author page:
a1805
,
Kiavash Movahedi
Kiavash Movahedi
Affiliation 1:
Myeloid Cell Immunology Laboratory, Brussels, Belgium
Affiliation 2:
Lab of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium
Bio-protocol author page:
a1806
and
Jo A Van Ginderachter
Jo A Van Ginderachter
Affiliation 1:
Myeloid Cell Immunology Laboratory, Brussels, Belgium
Affiliation 2:
Lab of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium
For correspondence:
jvangind@vub.ac.be
Bio-protocol author page:
a1807
,
date: 11/20/2014, 7734 views,
1 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1294
[Abstract] Tumors are heterogeneous microenvironments where complex interactions take place between neoplastic cells and infiltrating inflammatory cells, such as tumor-associated macrophages (TAM) and tumor-associated dendritic cells (TADC). The relevance of tumor-infiltrating ...
Article identifier: bio-protocol.org/e1294
Cell Biology
Analysis of Intestinal Permeability in Mice
Authors:
Jalaj Gupta
Jalaj Gupta
Affiliation:
Institute for Research in Biomedicine (IRB Barcelona), Barcelona, Spain
For correspondence:
Gupta@gsh.uni-frankfurt.de
Bio-protocol author page:
a1790
and
Angel R. Nebreda
Angel R. Nebreda
Affiliation 1:
Institute for Research in Biomedicine (IRB Barcelona), Barcelona, Spain
Affiliation 2:
Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain
For correspondence:
angel.nebreda@irbbarcelona.org
Bio-protocol author page:
a1791
,
date: 11/20/2014, 12553 views,
3 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1289
[Abstract] The intestinal epithelial layer serves as a barrier against pathogens and ingested toxins, which are present in the lumen of the intestine. The importance of the intestinal epithelial barrier is emphasized by the alterations in paracellular permeability ...
Article identifier: bio-protocol.org/e1289
Preparation of Golgi Membranes from Rat Liver
Authors:
Carmen Valente
Carmen Valente
Affiliation:
Institute of Protein Biochemistry (CNR), Naples, Italy
Bio-protocol author page:
a1792
,
Gabriele Turacchio
Gabriele Turacchio
Affiliation:
Institute of Protein Biochemistry (CNR), Naples, Italy
Bio-protocol author page:
a1793
,
Stefania Spanò
Stefania Spanò
Affiliation:
Institute of Protein Biochemistry (CNR), Naples, Italy
Bio-protocol author page:
a1794
,
Alberto Luini
Alberto Luini
Affiliation:
Institute of Protein Biochemistry (CNR), Naples, Italy
Bio-protocol author page:
a1795
and
Daniela Corda
Daniela Corda
Affiliation:
Institute of Protein Biochemistry (CNR), Naples, Italy
For correspondence:
d.corda@ibp.cnr.it
Bio-protocol author page:
a1796
,
date: 11/20/2014, 2391 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1291
[Abstract] This protocol details the isolation of enriched Golgi membranes from rat liver, using discontinuous density gradient centrifugation. This high-yield extraction method is useful for several applications, including immunoprecipitation of solubilised Golgi ...
Article identifier: bio-protocol.org/e1291
Separation of the Inner and Outer Mitochondrial Membrane in HeLa Cells
Authors:
Naotaka Nishimura
Naotaka Nishimura
Affiliation:
Department of Molecular Genetics, Kumamoto University, Kumamoto, Japan
Bio-protocol author page:
a1816
and
Masato Yano
Masato Yano
Affiliation:
Department of Medical Technology, Kumamoto Health Science University, Kumamoto, Japan
For correspondence:
yano@kumamoto-hsu.ac.jp
Bio-protocol author page:
a1817
,
date: 11/20/2014, 7607 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1299
[Abstract] Mitochondria are organelles that have important functions in oxidative phosphorylation, fatty acid oxidation and apoptosis signaling. They have two distinct membranes, outer membrane (OM) and inner membrane (IM). IM contains respiratory chain complexes ...
Article identifier: bio-protocol.org/e1299
Assay of Ornithine Decarboxylase and Spermidine/Spermine N
1
-acetyltransferase Activities
Authors:
Mervi T. Hyvönen
Mervi T. Hyvönen
Affiliation:
School of Pharmacy, University of Eastern Finland, Kuopio, Finland
For correspondence:
mervi.hyvonen@uef.fi
Bio-protocol author page:
a1820
,
Tuomo Keinänen
Tuomo Keinänen
Affiliation:
School of Pharmacy, University of Eastern Finland, Kuopio, Finland
Bio-protocol author page:
a1821
and
Leena Alhonen
Leena Alhonen
Affiliation:
School of Pharmacy, University of Eastern Finland, Kuopio, Finland
Bio-protocol author page:
a1822
,
date: 11/20/2014, 3082 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1301
[Abstract] The polyamines, spermidine (Spd) and spermine, and their diamine precursor putrescine, are important regulators of various cellular functions, such as proliferation and differentiation. Polyamine homeostasis is tightly regulated on the level of uptake, ...
Article identifier: bio-protocol.org/e1301
Microbiology
Purification and Sequencing of DNA Guides from Prokaryotic Argonaute
Authors:
Daan C. Swarts
Daan C. Swarts
Affiliation:
Department of Agrotechnology and Food Sciences, Wageningen University, Wageningen, the Netherlands
For correspondence:
daan.swarts@wur.nl
Bio-protocol author page:
a1688
,
Edze R. Westra
Edze R. Westra
Affiliation:
Department of Agrotechnology and Food Sciences, Wageningen University, Wageningen, the Netherlands
Bio-protocol author page:
a1801
,
Stan J. J. Brouns
Stan J. J. Brouns
Affiliation:
Department of Agrotechnology and Food Sciences, Wageningen University, Wageningen, the Netherlands
Bio-protocol author page:
a1802
and
John van der Oost
John van der Oost
Affiliation:
Department of Agrotechnology and Food Sciences, Wageningen University, Wageningen, the Netherlands
Bio-protocol author page:
a1690
,
date: 11/20/2014, 3429 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1293
[Abstract] Some proteins utilize nucleic acids to guide them to complementary nucleic acid targets. One example is prokaryotic Argonaute protein, which, binds small single stranded DNA molecules as guides (Swarts
et al.
, 2014). This protocol describes a method ...
Article identifier: bio-protocol.org/e1293
Infectious Focus Assays and Multiplicity of Infection (MOI) Calculations for Alpha-herpesviruses
Authors:
Anna Sloutskin
Anna Sloutskin
Affiliation:
Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel
Bio-protocol author page:
a1808
and
Ronald S. Goldstein
Ronald S. Goldstein
Affiliation:
Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel
For correspondence:
ron.goldstein@biu.ac.il
Bio-protocol author page:
a1809
,
date: 11/20/2014, 11608 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1295
[Abstract] Titration of viral stocks is a critical process before any experimental use of the virus. Here we describe an infectious focus assay for several alphaherpesviruses, a titration method for fluorescently labeled viruses, based on the original plaque assay. ...
Article identifier: bio-protocol.org/e1295
Semi-denaturing Detergent Agarose Gel Electrophoresis (SDD-AGE)
Authors:
Laura Molina-García
Laura Molina-García
Affiliation:
Department of Cellular and Molecular Biology, Centro de Investigaciones Biológicas-CSIC, Madrid, Spain
Bio-protocol author page:
a1812
and
Fátima Gasset-Rosa
Fátima Gasset-Rosa
Affiliation:
Department of Neurosciences, University of California, San Diego, USA
For correspondence:
fgasset@ucsd.edu
Bio-protocol author page:
a1813
,
date: 11/20/2014, 4511 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1297
[Abstract] Pathological proteins in neurodegenerative diseases suffer a conformational change to a misfolded amyloid state. Such pathological event leads to the aggregation of these proteins that indefinitely propagates as an altered form of itself, and harbor ...
Article identifier: bio-protocol.org/e1297
Competitive ELISA for Protein-lipopolysaccharide (LPS) Binding
Authors:
Victoria Martínez-Sernández
Victoria Martínez-Sernández
Affiliation:
Departamento de Microbiología y Parasitología, Universidad de Santiago de Compostela, Santiago de Compostela, Spain
Bio-protocol author page:
a1814
and
Florencio M. Ubeira
Florencio M. Ubeira
Affiliation:
Departamento de Microbiología y Parasitología, Universidad de Santiago de Compostela, Santiago de Compostela, Spain
For correspondence:
fm.ubeira@usc.es
Bio-protocol author page:
a1815
,
date: 11/20/2014, 3655 views,
1 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1298
[Abstract] Lipopolysaccharide is the major constituent of the outer membrane of gram-negative bacteria and, once released from the bacterial surface into the bloodstream, is a potent activator of the host immune system, which can lead to septic shock. LPS has a ...
Article identifier: bio-protocol.org/e1298
Isolation and Characterisation of Dendritic Cells from Peripheral Blood
Authors:
Matthew Reeves
Matthew Reeves
Affiliation:
Institute of Immunity and Transplantation, University College London, London, UK
For correspondence:
matthew.reeves@ucl.ac.uk
Bio-protocol author page:
a1818
and
John Sinclair
John Sinclair
Affiliation:
Institute of Immunity and Transplantation, University College London, London, UK
Bio-protocol author page:
a1819
,
date: 11/20/2014, 4890 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1300
[Abstract] Latency and reactivation of human cytomegalovirus (HCMV) is intimately associated with the myeloid lineage. Multiple studies have used
in vitro
protocols to generate dendritic cells (DCs) from myeloid precursors. Here we describe the direct isolation ...
Article identifier: bio-protocol.org/e1300
Secretion Assay in
Shigella flexneri
Authors:
Jonathan Reinhardt
Jonathan Reinhardt
Affiliation:
Max-Planck-Institute for Infection Biology, Berlin, Germany
Bio-protocol author page:
a1823
and
Michael Kolbe
Michael Kolbe
Affiliation:
Max-Planck-Institute for Infection Biology, Berlin, Germany
For correspondence:
kolbe@mpiib-berlin.mpg.de
Bio-protocol author page:
a1824
,
date: 11/20/2014, 3200 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1302
[Abstract]
Shigella flexneri
(
S. flexneri
) is a Gram-negative bacterium that causes gastroenteritis and shigellosis in humans. In order to establish and maintain an infection,
S. flexneri
utilises a type three secretion system (T3SS) to deliver virulence factors ...
Article identifier: bio-protocol.org/e1302
Plant Science
In vitro
Detection of S-acylation on Recombinant Proteins via the Biotin-Switch Technique
Authors:
Dong Qi
Dong Qi
Affiliation:
Department of Biology, Indiana University, Bloomington, USA
Present address:
Eurofins Lancaster Laboratory, Inc., Altria Client Services, Richmond, Virginia
Bio-protocol author page:
a1810
and
Roger W. Innes
Roger W. Innes
Affiliation:
Department of Biology, Indiana University, Bloomington, USA
For correspondence:
rinnes@indiana.edu
Bio-protocol author page:
a1811
,
date: 11/20/2014, 2707 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1296
[Abstract] Protein palmitoylation is the post-translational modification of proteins via the attachment of palmitate through acyl linkages. The nucleophile sulfhydryl group of cysteines is the common palmitoylation site. Covalent attachment of palmitate occurs ...
Article identifier: bio-protocol.org/e1296
Stem Cell
Isolation of FAP Cells from Mouse Dystrophic Skeletal Muscle Using Fluorescence Activated Cell Sorting
Authors:
Nicoletta Cordani
Nicoletta Cordani
Affiliation 1:
Scientific Institute, Lecco, Italy
Affiliation 2:
Department of Health of Science, University of Milano-Bicocca, Monza, Italy
Bio-protocol author page:
a1797
,
Viviana Pisa
Viviana Pisa
Affiliation:
Department of Biomedical and Clinical Sciences, University of Milano, Milan, Italy
Bio-protocol author page:
a1798
,
Laura Pozzi
Laura Pozzi
Affiliation:
Scientific Institute, Lecco, Italy
Bio-protocol author page:
a1799
and
Clara Sciorati
Clara Sciorati
Affiliation:
Division of Regenerative Medicine, Ospedale San Raffaele, Milan, Italy
For correspondence:
sciorati.clara@hsr.it
Bio-protocol author page:
a1800
,
date: 11/20/2014, 3160 views,
0 Q&A
DOI:
https://doi.org/10.21769/BioProtoc.1292
[Abstract] A population of muscle resident CD45
-
, CD31
-
cells expressing the mesenchymal PDGF receptor alpha (PDGFRα) as well as Sca-1 was first isolated in healthy mouse muscles in Uezumi
et al.
(2010). In the same year, Joe
et al.
(2010) identified and purified ...
Article identifier: bio-protocol.org/e1292
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