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Protocol for Murine/Mouse Platelets Isolation and Their Reintroduction in vivo

Featured protocol,  Authors: Jae Hong Im
Jae Hong ImAffiliation: CRUK-MRC Oxford Institute for Radiation Oncology, University of Oxford, Oxford, UK
Bio-protocol author page: a4021
 and Ruth J. Muschel
Ruth J. MuschelAffiliation: CRUK-MRC Oxford Institute for Radiation Oncology, University of Oxford, Oxford, UK
For correspondence: ruth.muschel@oncology.ox.ac.uk
Bio-protocol author page: a4022
date: 2/20/2017, 38 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2132.

Brief version appeared in Blood, Mar 2012
Platelets and coagulation have long been known to be essential for metastasis in experimental models. In order to study the interactions between tumor cells, platelets and endothelium, we have adapted methods used in coagulation research for the isolation of platelets and their reintroduction into mice. Anti-coagulated murine blood served as the source for platelets. Platelets were separated from other elements of the whole blood by centrifugation. Here the critical elements are first inhibition of coagulation and second isolation and maintenance of the platelets in the presence of inhibitors of platelet activation. We then used the vital dye PKH26 to fluorescently label the platelets. Infusion of these labelled platelets allows microscopic observation of the introduced platelets. After reintroduction, these platelets appear to function normally and comprise approximately 50% of the total platelets. Because they are fluorescently labelled, they can easily be identified. Finally it would be possible to use these methods for the determination of specific effects of altered gene expression in platelets by using platelets from genetically engineered mice. These methods have facilitated study of the interactions between platelets and tumor cells in tissue culture and in murine models. They would also be applicable to video microscopy. Here we provide details of the methods we have used for platelet isolation from mice and their staining for further microscopy and re-introduction into mice.

Analysis of the Virulence of Uropathogenic Escherichia coli Strain CFT073 in the Murine Urinary Tract

Featured protocol,  Authors: Anna Waldhuber
Anna WaldhuberAffiliation: Molekulare Pädiatrie, Dr. von Haunersches Kinderspital, Ludwig-Maximilians-Universität, München, Germany
Bio-protocol author page: a4062
Manoj Puthia
Manoj PuthiaAffiliation: Department of Microbiology, Immunology and Glycobiology (MIG), Institute of Laboratory Medicine, Lund University, Lund, Sweden
Bio-protocol author page: a4063
Andreas Wieser
Andreas WieserAffiliation: Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians-Universität, München, Germany
Bio-protocol author page: a4064
Catharina Svanborg
Catharina SvanborgAffiliation: Department of Microbiology, Immunology and Glycobiology (MIG), Institute of Laboratory Medicine, Lund University, Lund, Sweden
Bio-protocol author page: a4065
 and Thomas Miethke
Thomas Miethke Affiliation: Institute of Medical Microbiology and Hygiene, Medical Faculty of Mannheim, University of Heidelberg, Mannheim, Germany
For correspondence: thomas.miethke@medma.uni-heidelberg.de
Bio-protocol author page: a4056
date: 2/5/2017, 139 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2129.

Brief version appeared in J Clin Invest, Jul 2016
This urinary tract infection model was used to monitor the efficacy of a new virulence factor of the uropathogenic Escherichia coli strain CFT073 in vivo. The new virulence factor which we designated TIR-containing protein C (TcpC) blocks Toll-like receptor signaling and the NLRP3 inflammasome signaling cascade by interacting with key components of both pattern recognition receptor systems (Cirl et al., 2008; Waldhuber et al., 2016). We infected wild type and knock-out mice with wildtype CFT073 and a mutant CFT073 strain lacking tcpC. This protocol describes how the mice were infected, how CFT073 was prepared and how the infection was monitored. The protocol was derived from our previously published work and allowed us to demonstrate that TcpC is a powerful virulence factor by increasing the bacterial burden of CFT073 in the urine and kidneys. Moreover, TcpC was responsible for the development of kidney abscesses since infection of mice with wildtype but not tcpC-deficient CFT073 mutants caused this complication.

Protocol for Murine/Mouse Platelets Isolation and Their Reintroduction in vivo

Authors: Jae Hong Im
Jae Hong ImAffiliation: CRUK-MRC Oxford Institute for Radiation Oncology, University of Oxford, Oxford, UK
Bio-protocol author page: a4021
 and Ruth J. Muschel
Ruth J. MuschelAffiliation: CRUK-MRC Oxford Institute for Radiation Oncology, University of Oxford, Oxford, UK
For correspondence: ruth.muschel@oncology.ox.ac.uk
Bio-protocol author page: a4022
date: 2/20/2017, 38 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2132.

[Abstract] Platelets and coagulation have long been known to be essential for metastasis in experimental models. In order to study the interactions between tumor cells, platelets and endothelium, we have adapted methods used in coagulation research for the isolation of platelets and their reintroduction into mice. Anti-coagulated murine blood served as the source ...

Analysis of the Virulence of Uropathogenic Escherichia coli Strain CFT073 in the Murine Urinary Tract

Authors: Anna Waldhuber
Anna WaldhuberAffiliation: Molekulare Pädiatrie, Dr. von Haunersches Kinderspital, Ludwig-Maximilians-Universität, München, Germany
Bio-protocol author page: a4062
Manoj Puthia
Manoj PuthiaAffiliation: Department of Microbiology, Immunology and Glycobiology (MIG), Institute of Laboratory Medicine, Lund University, Lund, Sweden
Bio-protocol author page: a4063
Andreas Wieser
Andreas WieserAffiliation: Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians-Universität, München, Germany
Bio-protocol author page: a4064
Catharina Svanborg
Catharina SvanborgAffiliation: Department of Microbiology, Immunology and Glycobiology (MIG), Institute of Laboratory Medicine, Lund University, Lund, Sweden
Bio-protocol author page: a4065
 and Thomas Miethke
Thomas Miethke Affiliation: Institute of Medical Microbiology and Hygiene, Medical Faculty of Mannheim, University of Heidelberg, Mannheim, Germany
For correspondence: thomas.miethke@medma.uni-heidelberg.de
Bio-protocol author page: a4056
date: 2/5/2017, 139 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2129.

[Abstract] This urinary tract infection model was used to monitor the efficacy of a new virulence factor of the uropathogenic Escherichia coli strain CFT073 in vivo. The new virulence factor which we designated TIR-containing protein C (TcpC) blocks Toll-like receptor signaling and the NLRP3 inflammasome signaling cascade by interacting with key components of ...

Virus Binding and Internalization Assay for Adeno-associated Virus

Authors: Garrett E. Berry
Garrett E. BerryAffiliation 1: Gene Therapy Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA
Affiliation 2: Curriculum in Genetics and Molecular Biology, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA
For correspondence: berrygar@email.unc.edu
Bio-protocol author page: a3988
 and Longping V. Tse
Longping V. TseAffiliation: Gene Therapy Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA
Bio-protocol author page: a940
date: 1/20/2017, 176 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2110.

[Abstract] The binding and internalization of adeno-associated virus (AAV) is an important determinant of viral infectivity and tropism. The ability to dissect these two tightly connected cellular processes would allow better understanding and provide insight on virus entry and trafficking. In the following protocol, we describe a quantitative PCR (qPCR) based ...

Isolation of PBMCs Using Vacutainer® Cellular Preparation Tubes (CPTTM)

Authors: Alaina Puleo
Alaina PuleoAffiliation: Institute for Immunity, Transplantation, and Infection, Stanford University School of Medicine, Stanford, Palo Alto, CA, USA
Bio-protocol author page: a4007
Chantia Carroll
Chantia CarrollAffiliation: Institute for Immunity, Transplantation, and Infection, Stanford University School of Medicine, Stanford, Palo Alto, CA, USA
Bio-protocol author page: a4008
Holden Maecker
Holden MaeckerAffiliation: Institute for Immunity, Transplantation, and Infection, Stanford University School of Medicine, Stanford, Palo Alto, CA, USA
For correspondence: maecker@stanford.edu
Bio-protocol author page: a1861
 and Rohit Gupta
Rohit GuptaAffiliation: Institute for Immunity, Transplantation, and Infection, Stanford University School of Medicine, Stanford, Palo Alto, CA, USA
Bio-protocol author page: a1867
date: 1/20/2017, 274 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2103.

[Abstract] Peripheral blood mononuclear cell (PBMC) isolation is commonly done via density gradient centrifugation over Ficoll-Hypaque, a labor-intensive procedure that requires skilled technicians and can contribute to sample variability. Cellular Preparation Tubes (CPTs) are Vacutainer blood draw tubes that contain Ficoll-Hypaque and a gel plug that separates ...

Simultaneous Intranasal/Intravascular Antibody Labeling of CD4+ T Cells in Mouse Lungs

Authors: Yanqun Wang*
Yanqun WangAffiliation: State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
Bio-protocol author page: a3996
Jing Sun*
Jing SunAffiliation: State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
Bio-protocol author page: a3997
Rudragouda Channappanavar
Rudragouda ChannappanavarAffiliation: Departments of Microbiology, University of Iowa, Iowa, USA
Bio-protocol author page: a3999
Jingxian Zhao
Jingxian ZhaoAffiliation: State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
Bio-protocol author page: a3998
Stanley Perlman
Stanley PerlmanAffiliation: Departments of Microbiology, University of Iowa, Iowa, USA
For correspondence: stanley-perlman@uiowa.edu
Bio-protocol author page: a1251
 and Jincun Zhao
Jincun ZhaoAffiliation: State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou
For correspondence: zhaojincun@gird.cn
Bio-protocol author page: a1250
 (*contributed equally to this work) date: 1/5/2017, 277 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2099.

[Abstract] CD4+ T cell responses have been shown to be protective in many respiratory virus infections. In the respiratory tract, CD4+ T cells include cells in the airway and parenchyma and cells adhering to the pulmonary vasculature. Here we discuss in detail the methods that are useful for characterizing CD4+ T cells in different anatomic locations in mouse ...

In vitro Treatment of Mouse and Human Cells with Endogenous Ligands for Activation of the Aryl Hydrocarbon Receptor

Authors: Taisho Yamada
Taisho YamadaAffiliation: Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Molecular Medical Biochemistry Unit, Biological Chemistry and Engineering Course, Graduate School of Chemical Sciences and Engineering, Hokkaido University, Sapporo, Japan
Bio-protocol author page: a3976
 and Akinori Takaoka
Akinori TakaokaAffiliation: Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Molecular Medical Biochemistry Unit, Biological Chemistry and Engineering Course, Graduate School of Chemical Sciences and Engineering, Hokkaido University, Sapporo, Japan
For correspondence: takaoka@igm.hokudai.ac.jp
Bio-protocol author page: a2879
date: 1/5/2017, 219 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2097.

[Abstract] Activation of the aryl hydrocarbon receptor (AHR) by endogenous ligands has been implicated in a variety of physiological processes such as cell cycle regulation, cell differentiation and immune responses. It is reported that tryptophan metabolites, such as kynurenine (Kyn) and 6-formylindolo(3,2-b)carbazole (FICZ), are endogenous ligands for AHR (Stockinger ...

FICZ Exposure and Viral Infection in Mice

Authors: Taisho Yamada
Taisho YamadaAffiliation: Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Molecular Medical Biochemistry Unit, Biological Chemistry and Engineering Course, Graduate School of Chemical Sciences and Engineering, Hokkaido University, Sapporo, Japan
Bio-protocol author page: a3976
 and Akinori Takaoka
Akinori TakaokaAffiliation: Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Molecular Medical Biochemistry Unit, Biological Chemistry and Engineering Course, Graduate School of Chemical Sciences and Engineering, Hokkaido University, Sapporo, Japan
For correspondence: takaoka@igm.hokudai.ac.jp
Bio-protocol author page: a2879
date: 1/5/2017, 248 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2096.

[Abstract] The aryl hydrocarbon receptor (AHR) is known as a sensor for dioxins that mediates their toxicity, and also has important biophysiological roles such as circadian rhythms, cell differentiation and immune responses. 6-formylindolo(3,2-b)carbazole (FICZ), which is derived through the metabolism of L-tryptophan by ultraviolet B irradiation, is one of ...

Measuring Procaspase-8 and -10 Processing upon Apoptosis Induction

Authors: Sabine Pietkiewicz
Sabine PietkiewiczAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Present address: Medical advisory service of social health insurance, Essen, Germany
Bio-protocol author page: a3934
Clara Wolfe
Clara WolfeAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Bio-protocol author page: a3935
Jörn H. Buchbinder
Jörn H. BuchbinderAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Bio-protocol author page: a3936
 and Inna N. Lavrik
Inna N. LavrikAffiliation 1: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Affiliation 2: Federal research center Institute of Cytology and Genetics, Novosibirsk, Russia
For correspondence: inna.lavrik@med.ovgu.de
Bio-protocol author page: a3937
date: 1/5/2017, 284 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2081.

[Abstract] Apoptosis or programmed cell death is important for multicellular organisms to keep cell homeostasis and for the clearance of mutated or infected cells. Apoptosis can be induced by intrinsic or extrinsic stimuli. The first event in extrinsic apoptosis is the formation of the Death-Inducing Signalling Complex (DISC), where the initiator caspases-8 and ...

Quantitative Measurements of HIV-1 and Dextran Capture by Human Monocyte-derived Dendritic Cells (MDDCs)

Authors: Mickaël M. Ménager
Mickaël M. MénagerAffiliation: The Kimmel Center for Biology and Medicine of the Skirball Institute, New York University School of Medicine, New York, USA
For correspondence: mickael.menager@med.nyu.edu
Bio-protocol author page: a3703
 and Dan R. Littman
Dan R. LittmanAffiliation 1: The Kimmel Center for Biology and Medicine of the Skirball Institute, New York University School of Medicine, New York, USA
Affiliation 2: Howard Hughes Medical Institute, Washington, USA
Bio-protocol author page: a3704
date: 11/20/2016, 468 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2004.

[Abstract] The aim of this protocol is to describe how to measure and quantify the amount of HIV-1 particles and dextran molecules internalized in human monocyte derived dendritic cells (MDDCs), using three different techniques: flow cytometry, quantitative PCR and confocal microscopy....

Evaluation of Cross-presentation in Bone Marrow-derived Dendritic Cells in vitro and Splenic Dendritic Cells ex vivo Using Antigen-coated Beads

Authors: Andrés Alloatti
Andrés AlloattiAffiliation: Institute Curie, PSL Research University, INSERM U932, Paris, France
For correspondence: andres.alloatti@curie.fr
Bio-protocol author page: a3727
Fiorella Kotsias
Fiorella KotsiasAffiliation 1: Institute Curie, PSL Research University, INSERM U932, Paris, France
Affiliation 2: Department of Virology, University of Buenos Aires, Buenos Aires, Argentina
Bio-protocol author page: a3728
Eik Hoffmann
Eik HoffmannAffiliation 1: Institute Curie, PSL Research University, INSERM U932, Paris, France
Affiliation 2: VIB Inflammation Research Center, Unit of Molecular Signal Transduction in Inflammation, Ghent, Belgium
Affiliation 3: Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Bio-protocol author page: a3725
 and Sebastian Amigorena
Sebastian AmigorenaAffiliation: Institute Curie, PSL Research University, INSERM U932, Paris, France
Bio-protocol author page: a3730
date: 11/20/2016, 479 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2015.

[Abstract] Antigen presentation by MHC class I molecules, also referred to as cross-presentation, elicits cytotoxic immune responses. In particular, dendritic cells (DC) are the most proficient cross-presenting cells, since they have developed unique means to control phagocytic and degradative pathways.
   This protocol allows the evaluation of antigen cross-presentation ...
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In vitro Culture of Human PBMCs

Authors: Santosh K Panda
Santosh K PandaAffiliation: Infectious Disease Biology, Institute of Life Sciences, Bhubaneswar, India
Bio-protocol author page: a221
 and Balachandran Ravindran
Balachandran RavindranAffiliation: Infectious Disease Biology, Institute of Life Sciences, Bhubaneswar, India
For correspondence: ravindran8@gmail.com
Bio-protocol author page: a222
date: 2/5/2013, 39147 views, 4 Q&A
DOI: https://doi.org/10.21769/BioProtoc.322.

[Abstract] Peripheral blood mononuclear cells (PBMCs) consist of chiefly of lymphocytes and monocytes. Purified PBMCs are used in vitro to evaluate a variety of functions of lymphocytes viz; a) proliferation to mitogenic (PHA, Con-A) stimulation, b) monitoring of prior sensitisation in antigen recall assays by ...

Isolation and Culture of Mouse Bone Marrow-derived Macrophages (BMM’phi’)

Author: Ran Chen
Ran ChenAffiliation: Department of Genetics, Stanford University, Stanford, USA
For correspondence: rcchen@jfkbio.com
Bio-protocol author page: a34
date: 2/5/2012, 34381 views, 15 Q&A
DOI: https://doi.org/10.21769/BioProtoc.68.

[Abstract] Bone marrow derived macrophages are a type of white blood cell that can be isolated from mammalian bone marrow. In this protocol, a method is described in which bone marrow cells are isolated from mouse leg bones (femur and tibia), and then differentiated to bone marrow-derived macrophages in approximately ...

[Bio101] In vitro Differentiation of Mouse Th0, Th1 and Th2 from Naïve CD4 T Cells

Author: Jia Li
Jia LiAffiliation: Department of Immunology, Medical Center, Duke University, Durham, North Carolina, USA
For correspondence: jiali.email@gmail.com
Bio-protocol author page: a16
date: 11/20/2011, 29764 views, 18 Q&A
DOI: https://doi.org/10.21769/BioProtoc.157.

[Abstract] In vitro differentiation of helper T cells of various lineages is frequently used in T helper cell study. Naïve CD4 T cells can differentiate into certain lineage of T help cells in vitro in the presence of specific stimulatory cytokines and inhibition of cytokines that are essential for the differentiation ...

[Bio101] Whole Blood Staining of Human Monocyte Subsets for Flow Cytometry

Author: Zheng Liu
Zheng LiuAffiliation: The Feinstein Institute for Medical Research, Manhasset, NY, USA
For correspondence: zl2119@caa.columbia.edu
Bio-protocol author page: a12
Anne Davidson Lab, date: 5/20/2011, 21224 views, 10 Q&A
DOI: https://doi.org/10.21769/BioProtoc.69.

[Abstract] This is a general protocol to stain whole human blood for flow analysis with minimal spontaneous activation of monocytes. This protocol was developed or modified in Dr. Anne Davidson’s lab at Feinstein Institute for Medical Research....

[Bio101] In vivo BrdU Incorporation and Detection in Mouse

Author: Zheng Liu
Zheng LiuAffiliation: The Feinstein Institute for Medical Research, Manhasset, NY, USA
For correspondence: zl2119@caa.columbia.edu
Bio-protocol author page: a12
Anne Davidson Lab, date: 6/5/2011, 19025 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.81.

[Abstract] BrdU (Bromodeoxyuridine or 5-bromo-2’-deoxyuridine) is a synthetic nucleoside that is incorporated into DNA by proliferating cells. This protocol is to be used to incorporate and detect BrdU in murine plasma cells. The plasma cells described in this protocol are formed spontaneously in autoimmune mice ...

Isolation of Human PBMCs

Authors: Santosh K Panda
Santosh K PandaAffiliation: Infectious Disease Biology, Institute of Life Sciences, Bhubaneswar, India
Bio-protocol author page: a221
 and Balachandran Ravindran
Balachandran RavindranAffiliation: Infectious Disease Biology, Institute of Life Sciences, Bhubaneswar, India
For correspondence: ravindran8@gmail.com
Bio-protocol author page: a222
date: 2/5/2013, 18886 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.323.

[Abstract] Peripheral blood mononuclear cells (PBMCs) are chiefly lymphocytes and monocytes. PBMCs are separated from the whole blood by a density gradient centrifugation method using Ficoll Histopaque....

[Bio101] Thioglycollate Induced Peritonitis

Author: Zheng Liu
Zheng LiuAffiliation: The Feinstein Institute for Medical Research, Manhasset, NY, USA
For correspondence: zl2119@caa.columbia.edu
Bio-protocol author page: a12
Anne Davidson Lab, date: 6/20/2011, 17022 views, 3 Q&A
DOI: https://doi.org/10.21769/BioProtoc.84.

[Abstract] Intraperitoneal (i.p.) injection of thioglycollate elicits a robust influx of neutrophils into peritoneal cavity. The trafficking of the cells is believed to be mediated by chemokines CXCL1, CXCL2, and CXCL8 (Call et al., 2001; Cacalano et al., 1994). Thus this model can be used to test the ability ...

Intracellular Cytokine (INF-gamma) Staining Assay

Author: Huagang Zhang
Huagang ZhangAffiliation: Albert Einstein College of Medicine, Yeshiva University, New York City, USA
For correspondence: huagangzhang@gmail.com
Bio-protocol author page: a21
date: 4/5/2012, 16811 views, 2 Q&A
DOI: https://doi.org/10.21769/BioProtoc.122.

[Abstract] An intracellular cytokine (INF-gamma) staining assay is used to analyze the function of lymphocytes at the single cell level. By combining surface staining and intracellular cytokine staining, this assay can reveal the percentage of cytokine-releasing cells in a particular population, which cannot be ...

Isolation of Dendritic Cells and Macrophages from the Murine Kidneys of Lupus by Cell Sorter

Authors: Ramalingam Bethunaickan
Ramalingam BethunaickanAffiliation: Center for Autoimmunity and Musculoskeletal Diseases, Feinstein Institute for Medical Research, New York, USA
For correspondence: bramalingam@gmail.com
Bio-protocol author page: a24
 and Anne Davidson
Anne DavidsonAffiliation: Center for Autoimmunity and Musculoskeletal Diseases, Feinstein Institute for Medical Research, New York, USA
Bio-protocol author page: a1712
Anne Davidson Lab, date: 4/20/2012, 12428 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.168.

[Abstract] Methods for the isolation and characterization of mononuclear phagocytes from the kidneys of mice with SLE are essential to understand the patho-physiology of the disease. Activation of these cells is associated with the onset of clinical disease in mice and infiltration with these cells is associated ...

Bronchoalveolar Lavage and Lung Tissue Digestion

Authors: Hongwei Han
Hongwei HanAffiliation: Immunology Program, Benaroya Research Institute, Seattle, USA
For correspondence: hhan@benaroyaresearch.org
Bio-protocol author page: a544
 and Steven F. Ziegler
Steven F. ZieglerAffiliation: Immunology Program, Benaroya Research Institute, Seattle, USA
For correspondence: sziegler@benaroyaresearch.org
Bio-protocol author page: a543
date: 8/20/2013, 11627 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.859.

[Abstract] Bronchoalveolar lavage (BAL) is a simple but valuable and typically performed technique commonly used for studying the pathogenesis of lung diseases such as asthma and COPD. Cell counts can be combined with new methods for examining inflammatory responses, such as ELISA, Flow cytometric analysis, immunohistochemistry, ...
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