Mammalia

Tumorigenicity Assay in Nude Mice

Authors: Feng Du
Feng DuAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi'an, China
Bio-protocol author page: a4714
, Xiaodi Zhao
Xiaodi ZhaoAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi'an, China
For correspondence: leedyzhao@fmmu.edu.cn
Bio-protocol author page: a4715
and Daiming Fan
Daiming FanAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, China
Bio-protocol author page: a4716
, date: 7/5/2017, 120 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2364.

[Abstract] Tumorigenicity refers to the ability of cultured cells to develop viable tumors in immune-deficient animals. The goal of this protocol is to illustrate tumorigenicity assay by subcutaneous tumor-cell-transplantation in nude mice. Target cells are transplanted to 6-week-old nude mice subcutaneously and ...

Stereotaxic Surgery for Suprachiasmatic Nucleus Lesions in Mice

Authors: Kimiko Shimizu
Kimiko ShimizuAffiliation: Department of Biological Sciences, School of Science, the University of Tokyo, Tokyo, 113-0033 Japan
For correspondence: shimizuk@bs.s.u-tokyo.ac.jp
Bio-protocol author page: a4698
and Yoshitaka Fukada
Yoshitaka FukadaAffiliation: Department of Biological Sciences, School of Science, the University of Tokyo, Tokyo, 113-0033 Japan
Bio-protocol author page: a4699
, date: 6/20/2017, 198 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2346.

[Abstract] Site-specific lesions are invaluable methods for investigating the function of brain regions within the central nervous system and can be used to study neural mechanisms of behaviors. Precise stereotaxic surgery is required to lesion small regions of the brain such as the suprachiasmatic nucleus (SCN), ...

Olfactory Cued Learning Paradigm

Authors: Gary Liu
Gary LiuAffiliation 1: Program in Developmental Biology, Baylor College of Medicine, Houston, USA
Affiliation 2: Medical Scientist Training Program, Baylor College of Medicine, Houston, USA
For correspondence: garyliu87@gmail.com
Bio-protocol author page: a4435
, Cynthia K. McClard*
Cynthia K. McClardAffiliation 1: Medical Scientist Training Program, Baylor College of Medicine, Houston, USA
Affiliation 2: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, USA
Bio-protocol author page: a4436
, Burak Tepe*
Burak TepeAffiliation: Program in Developmental Biology, Baylor College of Medicine, Houston, USA
Bio-protocol author page: a4437
, Jessica Swanson
Jessica SwansonAffiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, USA
Bio-protocol author page: a4438
, Brandon Pekarek
Brandon PekarekAffiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, USA
Bio-protocol author page: a4439
, Sugi Panneerselvam
Sugi PanneerselvamAffiliation: Rice University, Houston, USA
Bio-protocol author page: a4440
and Benjamin R. Arenkiel
Benjamin R. ArenkielAffiliation 1: Program in Developmental Biology, Baylor College of Medicine, Houston, USA
Affiliation 2: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, USA
Affiliation 3: Department of Neuroscience, Baylor College of Medicine, Houston, USA
Affiliation 4: Jan and Dan Duncan Neurological Research Institute at Texas Children’s Hospital, Houston, USA
Bio-protocol author page: a4441
(*contributed equally to this work), date: 5/5/2017, 342 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2251.

[Abstract] Sensory stimulation leads to structural changes within the CNS (Central Nervous System), thus providing the fundamental mechanism for learning and memory. The olfactory circuit offers a unique model for studying experience-dependent plasticity, partly due to a continuous supply of integrating adult ...

Evaluation of Muscle Performance in Mice by Treadmill Exhaustion Test and Whole-limb Grip Strength Assay

Authors: Beatriz Castro
Beatriz CastroAffiliation: Department of Animal Sciences, Purdue University, West Lafayette, USA
For correspondence: beacb@purdue.edu
Bio-protocol author page: a4386
and Shihuan Kuang
Shihuan KuangAffiliation 1: Department of Animal Sciences, Purdue University, West Lafayette, USA
Affiliation 2: Center for Cancer Research, Purdue University, West Lafayette, USA
For correspondence: skuang@purdue.edu
Bio-protocol author page: a2572
, date: 4/20/2017, 528 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2237.

[Abstract] In vivo muscle function testing has become of great interest as primary phenotypic analysis of muscle performance. This protocol provides detailed procedures to perform the treadmill exhaustion test and the whole-limb grip strength assay, two methods commonly used in the neuromuscular research field....

The Object Context-place-location Paradigm for Testing Spatial Memory in Mice

Authors: Edith Lesburguères
Edith LesburguèresAffiliation: Center for Neural Science, New York University, New York, USA
Bio-protocol author page: a4246
, Panayiotis Tsokas
Panayiotis TsokasAffiliation 1: Department of Physiology and Pharmacology, the Robert F. Furchgott Center for Neural and Behavioral Science, Downstate Medical Center, State University of New York, New York, USA
Affiliation 2: Department of Anesthesiology, the Robert F. Furchgott Center for Neural and Behavioral Science, Downstate Medical Center, State University of New York, New York, USA
Bio-protocol author page: a4243
, Todd Charlton Sacktor
Todd Charlton SacktorAffiliation 1: Department of Physiology and Pharmacology, the Robert F. Furchgott Center for Neural and Behavioral Science, Downstate Medical Center, State University of New York, New York, USA
Affiliation 2: Department of Neurology, the Robert F. Furchgott Center for Neural and Behavioral Science, Downstate Medical Center, State University of New York, New York, USA
Bio-protocol author page: a4244
and André Antonio Fenton
André Antonio FentonAffiliation 1: Center for Neural Science, New York University, New York, USA
Affiliation 2: Department of Physiology and Pharmacology, the Robert F. Furchgott Center for Neural and Behavioral Science, Downstate Medical Center, State University of New York, New York, USA
For correspondence: afenton@nyu.edu
Bio-protocol author page: a4245
, date: 4/20/2017, 525 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2231.

[Abstract] This protocol was originally designed to examine long-term spatial memory in PKMζ knockout (i.e., PKMζ-null) mice (Tsokas et al., 2016). Our main goal was to test whether the ability of these animals to maintain previously acquired spatial information was sensitive to the type and complexity of the ...

Measurement of Dipeptidylpeptidase Activity in vitro and in vivo

Authors: Rosa Barreira da Silva
Rosa Barreira da SilvaAffiliation: Cancer Immunology, Genentech, South San Francisco, CA, USA
Bio-protocol author page: a4161
, Molly A. Ingersoll
Molly A. IngersollAffiliation 1: Laboratory of Dendritic Cell Immunobiology, Institut Pasteur, Paris, France
Affiliation 2: INSERM U1223, Paris, France
Bio-protocol author page: a4162
and Matthew L. Albert
Matthew L. AlbertAffiliation 1: Cancer Immunology, Genentech, South San Francisco, CA, USA
Affiliation 2: Laboratory of Dendritic Cell Immunobiology, Institut Pasteur, Paris, France
Affiliation 3: INSERM U1223, Paris, France
For correspondence: albertm7@gene.com
Bio-protocol author page: a4163
, date: 3/20/2017, 561 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2184.

[Abstract] Dipeptidylpeptidases (DPPs) are serine proteases, which cleave small proteins and peptides possessing a proline or an alanine in the second position of their N-terminus. Among the members of this family, dipeptidylpeptidase 4 (DPP4) is constitutively expressed in the extracellular space. DPP4 is found ...

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, 930 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 ...

Preparation of Purified Gram-positive Bacterial Cell Wall and Detection in Placenta and Fetal Tissues

Authors: Beth Mann
Beth MannAffiliation: Department of Infectious Diseases, St Jude Children’s Research Hospital, Memphis TN, USA
Bio-protocol author page: a3796
, Lip Nam Loh
Lip Nam LohAffiliation: Department of Infectious Diseases, St Jude Children’s Research Hospital, Memphis TN, USA
Bio-protocol author page: a3797
, Geli Gao
Geli GaoAffiliation: Department of Infectious Diseases, St Jude Children’s Research Hospital, Memphis TN, USA
Bio-protocol author page: a3798
and Elaine Tuomanen
Elaine TuomanenAffiliation: Department of Infectious Diseases, St Jude Children’s Research Hospital, Memphis TN, USA
For correspondence: Elaine.tuomanen@stjude.org
Bio-protocol author page: a3799
, date: 12/5/2016, 1265 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2037.

[Abstract] Cell wall is a complex biopolymer on the surface of all Gram-positive bacteria. During infection, cell wall is recognized by the innate immune receptor Toll-like receptor 2 causing intense inflammation and tissue damage. In animal models, cell wall traffics from the blood stream to many organs in the ...

Delayed Spatial Win-shift Test on Radial Arm Maze

Authors: Simone N. De Luca
Simone N. De LucaAffiliation: School of Health and Biomedical Sciences, RMIT University, Melbourne, Vic, Australia
Bio-protocol author page: a3841
, Luba Sominsky
Luba Sominsky Affiliation: School of Health and Biomedical Sciences, RMIT University, Melbourne, Vic, Australia
Bio-protocol author page: a3842
and Sarah J. Spencer
Sarah J. SpencerAffiliation: School of Health and Biomedical Sciences, RMIT University, Melbourne, Vic, Australia
For correspondence: Sarah.Spencer@rmit.edu.au
Bio-protocol author page: a3843
, date: 12/5/2016, 1145 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2053.

[Abstract] The radial arm maze (RAM) is used to assess reference and working memory in rodents. This task relies on the rodent’s ability to orientate itself in the maze using extra-maze visual cues. This test can be used to investigate whether a rodent’s cognition is improved or impaired under a variety of experimental ...

Identification of Methylated Deoxyadenosines in Genomic DNA by dA^6m DNA Immunoprecipitation

Authors: Magdalena J. Koziol
Magdalena J. KoziolAffiliation 1: Wellcome Trust Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge, UK
Affiliation 2: Department of Zoology, University of Cambridge, Cambridge, UK
For correspondence: mjk39@cam.ac.uk
Bio-protocol author page: a3666
, Charles R. Bradshaw
Charles R. BradshawAffiliation: Wellcome Trust Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge, UK
Bio-protocol author page: a3667
, George E. Allen
George E. AllenAffiliation: Wellcome Trust Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge, UK
Bio-protocol author page: a3668
, Ana S. H. Costa
Ana S. H. CostaAffiliation: Medical Research Council Cancer Unit, University of Cambridge, Hutchison/MRC Research Centre, Cambridge, UK
Bio-protocol author page: a3669
and Christian Frezza
Christian FrezzaAffiliation: Medical Research Council Cancer Unit, University of Cambridge, Hutchison/MRC Research Centre, Cambridge, UK
Bio-protocol author page: a3670
, date: 11/5/2016, 1143 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1990.

[Abstract] dA^6m DNA immunoprecipitation followed by deep sequencing (DIP-Seq) is a key tool in identifying and studying the genome-wide distribution of N⁶-methyldeoxyadenosine (dA^6m). The precise function of this novel DNA modification remains to be fully elucidated, but it is known to be absent from transcriptional ...

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[Bio101] Glucose Tolerance Test in Mice

Author: Peichuan Zhang
Peichuan ZhangAffiliation 1: Department of Biology, The Pennsylvania State University, University Park, PA, USA
Affiliation 2: Department of Biochemistry and Biophysics, University of California, San Francisco, USA
For correspondence: peichuan.zhang@ucsf.edu
Bio-protocol author page: a11
, date: 10/5/2011, 32040 views, 4 Q&A
DOI: https://doi.org/10.21769/BioProtoc.159.

[Abstract] Glucose tolerance test is a standard procedure that addresses how quickly exogenous glucose can be cleared from blood. Specifically, uptake of glucose from the blood by cells is regulated by insulin. Impairment of glucose tolerance (i.e, longer time to clear given amount of glucose) indicates problems ...

[Bio101] Subcutaneous Injection of Tumor Cells

Author: Jason Reuter, date: 12/20/2011, 26663 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.166.

[Abstract] Growth of cells in the subcutaneous space of immunocompromised mice is a common method for assaying tumorigenic potential in vivo. This technique is also used to assess the effects of therapeutic interventions on cancer cell lines....

In vivo Matrigel Plug Angiogenesis Assay

Authors: Hong Lok Lung
Hong Lok LungAffiliation: Department of Clinical Oncology and Center for Cancer Research, The University of Hong Kong, Hong Kong , Hong Kong SAR
Bio-protocol author page: a102
and Maria Li Lung
Maria Li LungAffiliation: Department of Clinical Oncology and Center for Cancer Research, The University of Hong Kong, Hong Kong , Hong Kong SAR
For correspondence: mlilung@hku.hk
Bio-protocol author page: a101
, date: 9/20/2012, 24667 views, 2 Q&A
DOI: https://doi.org/10.21769/BioProtoc.261.

[Abstract] The matrigel plug angiogenesis assay is a simple in vivo technique to detect the newly formed blood vessels in the transplanted gel plugs in nude mice. The matrigel matrix is derived from the engelbroth-holm-swarm (EHS) mouse sarcoma, and its composition is comparable to the basement membrane proteins. ...

[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, 18726 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 ...

Alcian Blue – Alizarin Red Staining of Mouse Skeleton

Author: Peichuan Zhang
Peichuan ZhangAffiliation 1: Department of Biology, The Pennsylvania State University, University Park, PA, USA
Affiliation 2: Department of Biochemistry and Biophysics, University of California, San Francisco, CA, USA
For correspondence: peichuan.zhang@ucsf.edu
Bio-protocol author page: a11
, date: 4/20/2012, 16606 views, 9 Q&A
DOI: https://doi.org/10.21769/BioProtoc.162.

[Abstract] Our lab has used the Alcian blue – Alizarin red staining method with certain modifications to characterize skeleton deformities in mice lacking Pek/Perk, encoding a translational control eIF2alpha kinase....

A Quick, No Frills Approach to Mouse Genotyping

Author: Manuel E. Lopez
Manuel E. LopezAffiliation: Department of Developmental Biology, Stanford University, Stanford, USA
For correspondence: manduardo@gmail.com
Bio-protocol author page: a74
, date: 8/5/2012, 12325 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.244.

[Abstract] Mice are extremely powerful mammalian genetic model organisms for basic and medical research, but managing a colony of transgenic mice is time consuming and expensive, many times requiring the help of dedicated technicians. Slow and laborious genotyping procedures add to the hassle. Outsourcing is costly ...

Iron - Prussian Blue Reaction - Mallory’s Method

Author: Hani Jouihan
Hani JouihanAffiliation: Division of Endocrinology, Gerontology and Metabolism, Stanford University School of Medicine, Stanford, USA
For correspondence: hanijouihan@hotmail.com
Bio-protocol author page: a52
, date: 7/5/2012, 11696 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.222.

[Abstract] Purpose: To demonstrate ferric iron in tissue sections. Small amounts of iron are found normally in spleen and bone marrow. Excessive amounts are present in hemochromatosis, with deposits found in the liver and pancreas, hemosiderosis, with deposits in the liver, spleen, and lymph nodes.
Principle: The ...

[Bio101] Isolation of Endothelial Cells from Mice

Author: Huan Pang
Huan PangAffiliation: Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, USA
For correspondence: pang_huan@hotmail.com
Bio-protocol author page: a48
, date: 4/20/2012, 11592 views, 4 Q&A
DOI: https://doi.org/10.21769/BioProtoc.205.

[Abstract] Endothelial cells line the entire circulatory system, from the heart to the smallest capillary. In this protocol, attaining and maintaining vitality of the organ is critical. Minimizing the processing time between removing the organ and beginning the digestions will improve yields as cells are more ...

[Bio101] Chromatin Immunoprecipitation (ChIP) Protocol from Tissue

Author: Nabila Aboulaich, date: 1/20/2011, 10773 views, 2 Q&A
DOI: https://doi.org/10.21769/BioProtoc.21.

[Abstract] ...

[Bio101] Murine xenograft model

Author: FengZhi Liu
FengZhi LiuAffiliation: School of Biomedical Sciences, Thomas Jefferson University, Philadelphia, USA
For correspondence: fengzhi6@yahoo.com
Bio-protocol author page: a51
, date: 6/5/2012, 10248 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.221.

[Abstract] This experiment is used to test the efficacies of chemo treatments or gene therapy in an in vivo system. In this protocol, the mouse xenograft model is used....

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Mammalia

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Author: Jason Reuter, date: 12/20/2011, 26663 views, 1 Q&ADOI: https://doi.org/10.21769/BioProtoc.166.

Author: Zheng Liu Zheng LiuAffiliation: The Feinstein Institute for Medical Research, Manhasset, NY, USAFor correspondence: zl2119@caa.columbia.eduBio-protocol author page: a12, Anne Davidson Lab, date: 6/20/2011, 18726 views, 3 Q&ADOI: https://doi.org/10.21769/BioProtoc.84.

Author: Manuel E. Lopez Manuel E. LopezAffiliation: Department of Developmental Biology, Stanford University, Stanford, USAFor correspondence: manduardo@gmail.comBio-protocol author page: a74, date: 8/5/2012, 12325 views, 1 Q&ADOI: https://doi.org/10.21769/BioProtoc.244.

Author: Huan Pang Huan PangAffiliation: Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, USAFor correspondence: pang_huan@hotmail.comBio-protocol author page: a48, date: 4/20/2012, 11592 views, 4 Q&ADOI: https://doi.org/10.21769/BioProtoc.205.

Author: Nabila Aboulaich, date: 1/20/2011, 10773 views, 2 Q&ADOI: https://doi.org/10.21769/BioProtoc.21.