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Differential Salt Fractionation of Nuclei to Analyze Chromatin-associated Proteins from Cultured Mammalian Cells

Featured protocol,  Authors: Christin Herrmann
Christin HerrmannAffiliation 1: Division of Cancer Pathobiology, Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia, Philadelphia, USA
Affiliation 2: Cell & Molecular Biology Graduate Group, University of Pennsylvania, Philadelphia, USA
Bio-protocol author page: a4254
Daphne C. Avgousti
Daphne C. AvgoustiAffiliation 1: Division of Cancer Pathobiology, Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia, Philadelphia, USA
Affiliation 2: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, USA
Bio-protocol author page: a4255
 and Matthew D. Weitzman
Matthew D. WeitzmanAffiliation 1: Division of Cancer Pathobiology, Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia, Philadelphia, USA
Affiliation 2: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, USA
For correspondence: weitzmanm@email.chop.edu
Bio-protocol author page: a4256
date: 3/20/2017, 123 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2175.

Brief version appeared in Nature, Jul 2016
Nucleosomes are the core units of cellular chromatin and are comprised of 147 base pairs (bp) of DNA wrapped around an octamer of histone proteins. Proteins such as chromatin remodelers, transcription factors, and DNA repair proteins interact dynamically with chromatin to regulate access to DNA, control gene transcription, and maintain genome integrity. The extent of association with chromatin changes rapidly in response to stresses, such as immune activation, oxidative stress, or viral infection, resulting in downstream effects on chromatin conformation and transcription of target genes. To elucidate changes in the composition of proteins associated with chromatin under different conditions, we adapted existing protocols to isolate nuclei and fractionate cellular chromatin using a gradient of salt concentrations. The presence of specific proteins in different salt fractions can be assessed by Western blotting or mass spectrometry, providing insight into the degree to which they are associated with chromatin.

RNA-protein UV-crosslinking Assay

Featured protocol,  Authors: Dipak Kumar Poria
Dipak Kumar PoriaAffiliation: Department of Biological Sciences, Indian Institute of Science Education and Research, Kolkata, India
Bio-protocol author page: a4215
 and Partho Sarothi Ray
Partho Sarothi RayAffiliation: Department of Biological Sciences, Indian Institute of Science Education and Research, Kolkata, India
For correspondence: psray@iiserkol.ac.in
Bio-protocol author page: a4216
date: 3/20/2017, 176 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2193.

Brief version appeared in Oncogene, Mar 2016
RNA-protein interactions play a crucial role in every aspect of RNA metabolism, and also plays a major role in post-transcriptional gene regulation. RNA-binding proteins have been implicated in viral gene expression (Ray and Das, 2002) and microRNA-mediated gene regulation (Poria et al., 2016). Here we have described the protocol which (1) covalently links transiently interacting RNA-protein complexes by UV crosslinking, (2) removes the unprotected RNA by RNase digestion and (3) detects the RNA-protein complexes by SDS-PAGE analysis. This protocol provides a rapid and reliable means to directly assay RNA-protein interactions and their kinetics using purified proteins and also help in identifying novel RNA-protein interactions

Polysome Analysis

Featured protocol,  Authors: Dipak Kumar Poria
Dipak Kumar PoriaAffiliation: Department of Biological Sciences, Indian Institute of Science Education and Research, Kolkata, India
Bio-protocol author page: a4215
 and Partho Sarothi Ray
Partho Sarothi RayAffiliation: Department of Biological Sciences, Indian Institute of Science Education and Research, Kolkata, India
For correspondence: psray@iiserkol.ac.in
Bio-protocol author page: a4216
date: 3/20/2017, 144 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2192.

Brief version appeared in Oncogene, Mar 2016
Polysome analysis is a method to separate mRNAs from a cell into actively translating and non-translating fractions depending on their association with polysomes. By this protocol, cell lysates are fractionated by sucrose density gradient ultracentrifugation. Free mRNA fraction and various ribosomal fractions, such as 40S, 60S, monosomes and polysomes are collected by fractionation. Association of particular mRNAs with these fractions is detected by reverse transcription – PCR to investigate the translational state of the mRNA.

3D Stroma Invasion Assay

Featured protocol,  Authors: Yvette May Coulson-Thomas
Yvette May Coulson-ThomasAffiliation: Department of Biochemistry, Universidade Federal de São Paulo, São Paulo, Brazil
For correspondence: ycoulsonthomas@gmail.com
Bio-protocol author page: a4214
 and Vivien Jane Coulson-Thomas
Vivien Jane Coulson-ThomasAffiliation: College of Optometry, the Ocular Surface Institute (TOSI), University of Houston, Houston, USA
For correspondence: vcoulsonthomas@gmail.com
Bio-protocol author page: a1653
date: 3/20/2017, 99 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2195.

Brief version appeared in Cell Tissue Res, Nov 2011
We have developed a 3D co-culture system composed of fibroblasts and colorectal cancer cells that enables us to study the desmoplastic reaction. This method also enables us to study the influence of the desmoplastic reaction on the migration of colorectal cancer cells through the surrounding stroma. This protocol has been previously published (Coulson-Thomas et al., 2011) and is described here in more detail.

Ca2+ Measurements in Mammalian Cells with Aequorin-based Probes

Featured protocol,  Authors: Anna Tosatto
Anna TosattoAffiliation: Department of Biomedical Sciences, University of Padua, Padua, Italy
Bio-protocol author page: a4128
Rosario Rizzuto
Rosario RizzutoAffiliation: Department of Biomedical Sciences, University of Padua, Padua, Italy
For correspondence: rosario.rizzuto@unipd.it
Bio-protocol author page: a4129
 and Cristina Mammucari
Cristina MammucariAffiliation: Department of Biomedical Sciences, University of Padua, Padua, Italy
For correspondence: cristina.mammucari@unipd.it
Bio-protocol author page: a4130
date: 3/5/2017, 213 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2155.

Brief version appeared in EMBO Mol Med, May 2016
Aequorin is a Ca2+ sensitive photoprotein suitable to measure intracellular Ca2+ transients in mammalian cells. Thanks to recombinant cDNAs expression, aequorin can be specifically targeted to various subcellular compartments, thus allowing an accurate measurement of Ca2+ uptake and release of different intracellular organelles. Here, we describe how to use this probe to measure cytosolic Ca2+ levels and mitochondrial Ca2+ uptake in mammalian cells.

Liposome Flotation Assays for Phosphoinositide-protein Interaction

Featured protocol,  Authors: Helene Tronchere
Helene TronchereAffiliation: INSERM U1048 I2MC and Université Paul Sabatier, Toulouse, France
Bio-protocol author page: a4209
 and Frederic Boal
Frederic BoalAffiliation: INSERM U1048 I2MC and Université Paul Sabatier, Toulouse, France
For correspondence: frederic.boal@inserm.fr
Bio-protocol author page: a4210
date: 3/5/2017, 208 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2169.

Brief version appeared in J Cell Sci, Feb 2015
Phosphoinositides are rare membrane lipids involved in the control of the major cellular functions and signaling pathways. They are able to recruit specific effector proteins to the cytosolic face of plasma membrane and organelles to coordinate a vast variety of signaling and trafficking processes, as well to maintain specific identity of the different subcellular compartments (Di Paolo and De Camilli, 2006; Lemmon, 2003). Therefore, analysis of these effectors’ binding properties and specificity towards different phosphoinositides is crucial for the understanding of their cellular functions. This protocol describes a method to characterize the binding of proteins to different phosphoinositide-containing vesicles.

Differential Salt Fractionation of Nuclei to Analyze Chromatin-associated Proteins from Cultured Mammalian Cells

Authors: Christin Herrmann
Christin HerrmannAffiliation 1: Division of Cancer Pathobiology, Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia, Philadelphia, USA
Affiliation 2: Cell & Molecular Biology Graduate Group, University of Pennsylvania, Philadelphia, USA
Bio-protocol author page: a4254
Daphne C. Avgousti
Daphne C. AvgoustiAffiliation 1: Division of Cancer Pathobiology, Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia, Philadelphia, USA
Affiliation 2: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, USA
Bio-protocol author page: a4255
 and Matthew D. Weitzman
Matthew D. WeitzmanAffiliation 1: Division of Cancer Pathobiology, Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia, Philadelphia, USA
Affiliation 2: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, USA
For correspondence: weitzmanm@email.chop.edu
Bio-protocol author page: a4256
date: 3/20/2017, 123 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2175.

[Abstract] Nucleosomes are the core units of cellular chromatin and are comprised of 147 base pairs (bp) of DNA wrapped around an octamer of histone proteins. Proteins such as chromatin remodelers, transcription factors, and DNA repair proteins interact dynamically with chromatin to regulate access to DNA, control gene transcription, and maintain genome integrity. ...

RNA-protein UV-crosslinking Assay

Authors: Dipak Kumar Poria
Dipak Kumar PoriaAffiliation: Department of Biological Sciences, Indian Institute of Science Education and Research, Kolkata, India
Bio-protocol author page: a4215
 and Partho Sarothi Ray
Partho Sarothi RayAffiliation: Department of Biological Sciences, Indian Institute of Science Education and Research, Kolkata, India
For correspondence: psray@iiserkol.ac.in
Bio-protocol author page: a4216
date: 3/20/2017, 176 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2193.

[Abstract] RNA-protein interactions play a crucial role in every aspect of RNA metabolism, and also plays a major role in post-transcriptional gene regulation. RNA-binding proteins have been implicated in viral gene expression (Ray and Das, 2002) and microRNA-mediated gene regulation (Poria et al., 2016). Here we have described the protocol which (1) covalently ...

Polysome Analysis

Authors: Dipak Kumar Poria
Dipak Kumar PoriaAffiliation: Department of Biological Sciences, Indian Institute of Science Education and Research, Kolkata, India
Bio-protocol author page: a4215
 and Partho Sarothi Ray
Partho Sarothi RayAffiliation: Department of Biological Sciences, Indian Institute of Science Education and Research, Kolkata, India
For correspondence: psray@iiserkol.ac.in
Bio-protocol author page: a4216
date: 3/20/2017, 144 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2192.

[Abstract] Polysome analysis is a method to separate mRNAs from a cell into actively translating and non-translating fractions depending on their association with polysomes. By this protocol, cell lysates are fractionated by sucrose density gradient ultracentrifugation. Free mRNA fraction and various ribosomal fractions, such as 40S, 60S, monosomes and polysomes ...

3D Stroma Invasion Assay

Authors: Yvette May Coulson-Thomas
Yvette May Coulson-ThomasAffiliation: Department of Biochemistry, Universidade Federal de São Paulo, São Paulo, Brazil
For correspondence: ycoulsonthomas@gmail.com
Bio-protocol author page: a4214
 and Vivien Jane Coulson-Thomas
Vivien Jane Coulson-ThomasAffiliation: College of Optometry, the Ocular Surface Institute (TOSI), University of Houston, Houston, USA
For correspondence: vcoulsonthomas@gmail.com
Bio-protocol author page: a1653
date: 3/20/2017, 99 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2195.

[Abstract] We have developed a 3D co-culture system composed of fibroblasts and colorectal cancer cells that enables us to study the desmoplastic reaction. This method also enables us to study the influence of the desmoplastic reaction on the migration of colorectal cancer cells through the surrounding stroma. This protocol has been previously published (Coulson-Thomas ...

Ca2+ Measurements in Mammalian Cells with Aequorin-based Probes

Authors: Anna Tosatto
Anna TosattoAffiliation: Department of Biomedical Sciences, University of Padua, Padua, Italy
Bio-protocol author page: a4128
Rosario Rizzuto
Rosario RizzutoAffiliation: Department of Biomedical Sciences, University of Padua, Padua, Italy
For correspondence: rosario.rizzuto@unipd.it
Bio-protocol author page: a4129
 and Cristina Mammucari
Cristina MammucariAffiliation: Department of Biomedical Sciences, University of Padua, Padua, Italy
For correspondence: cristina.mammucari@unipd.it
Bio-protocol author page: a4130
date: 3/5/2017, 213 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2155.

[Abstract] Aequorin is a Ca2+ sensitive photoprotein suitable to measure intracellular Ca2+ transients in mammalian cells. Thanks to recombinant cDNAs expression, aequorin can be specifically targeted to various subcellular compartments, thus allowing an accurate measurement of Ca2+ uptake and release of different intracellular organelles. Here, we describe how ...

Liposome Flotation Assays for Phosphoinositide-protein Interaction

Authors: Helene Tronchere
Helene TronchereAffiliation: INSERM U1048 I2MC and Université Paul Sabatier, Toulouse, France
Bio-protocol author page: a4209
 and Frederic Boal
Frederic BoalAffiliation: INSERM U1048 I2MC and Université Paul Sabatier, Toulouse, France
For correspondence: frederic.boal@inserm.fr
Bio-protocol author page: a4210
date: 3/5/2017, 208 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2169.

[Abstract] Phosphoinositides are rare membrane lipids involved in the control of the major cellular functions and signaling pathways. They are able to recruit specific effector proteins to the cytosolic face of plasma membrane and organelles to coordinate a vast variety of signaling and trafficking processes, as well to maintain specific identity of the different ...

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, 345 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 ...

A Murine Orthotopic Allograft to Model Prostate Cancer Growth and Metastasis

Authors: Robert M. Hughes
Robert M. HughesAffiliation 1: The James Buchanan Brady Urological Institute, Department of Urology, Johns Hopkins School of Medicine, Baltimore, USA
Affiliation 2: The Department of Oncology, Johns Hopkins School of Medicine, Baltimore, USA
Affiliation 3: The Sidney Kimmel Cancer Center, Johns Hopkins School of Medicine, Baltimore, USA
Bio-protocol author page: a4138
Brian W. Simons
Brian W. SimonsAffiliation: The James Buchanan Brady Urological Institute, Department of Urology, Johns Hopkins School of Medicine, Baltimore, USA
Bio-protocol author page: a4139
 and Paula J. Hurley
Paula J. HurleyAffiliation 1: The James Buchanan Brady Urological Institute, Department of Urology, Johns Hopkins School of Medicine, Baltimore, USA
Affiliation 2: The Department of Oncology, Johns Hopkins School of Medicine, Baltimore, USA
Affiliation 3: The Sidney Kimmel Cancer Center, Johns Hopkins School of Medicine, Baltimore, USA
For correspondence: phurley2@jhmi.edu
Bio-protocol author page: a4140
date: 2/20/2017, 312 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2137.

[Abstract] Prostate cancer is one of the most common cancers in men in the United States. Comprehensive understanding of the biology contributing to prostate cancer will have important clinical implications. Animal models have greatly impacted our knowledge of disease and will continue to be a valuable resource for future studies. Herein, we describe a detailed ...

In situ Hybridization (ISH) and Quantum Dots (QD) of miRNAs

Authors: Sajni Josson
Sajni JossonAffiliation: Uro-Oncology Research Program, Department of Medicine, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, USA
Present address: Neostrata Inc, Princeton, USA
For correspondence: sajnij@gmail.com
Bio-protocol author page: a4072
Murali Gururajan
Murali GururajanAffiliation: Uro-Oncology Research Program, Department of Medicine, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, USA
Present address: Bristol-Myers Squibb Inc, Princeton, USA
For correspondence: gururajanmurali@gmail.com
Bio-protocol author page: a4073
 and Leland W.K. Chung
Leland W.K. ChungAffiliation: Uro-Oncology Research Program, Department of Medicine, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, USA
For correspondence: leland.chung@cshs.org
Bio-protocol author page: a4074
date: 2/20/2017, 328 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2138.

[Abstract] miRNA are short non-coding RNA which inhibit translation of mRNA. miRNA regulate several cellular processes. Certain miRNA are known to induce oncogenesis. miRNA can be measured by real-time PCR and be imaged using a combination of in situ hybridization (ISH) and quantum dots (QD). The advantage of using quantum dots is that several miRNA can be simultaneously ...

miRNA Characterization from the Extracellular Vesicles

Authors: Sajni Josson
Sajni JossonAffiliation 1: Uro-Oncology Research Program, Department of Medicine, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, USA
Affiliation 2: Neostrata Inc, Princeton, USA
For correspondence: sajni.j@gmail.com
Bio-protocol author page: a4072
Murali Gururajan
Murali GururajanAffiliation 1: Uro-Oncology Research Program, Department of Medicine, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, USA
Affiliation 2: Bristol-Myers Squibb Inc, Princeton, USA
For correspondence: gururajanmurali@gmail.com
Bio-protocol author page: a4073
 and Leland W.K. Chung
Leland W.K. ChungAffiliation: Uro-Oncology Research Program, Department of Medicine, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, USA
For correspondence: leland.chung@cshs.org
Bio-protocol author page: a4074
date: 2/20/2017, 265 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2139.

[Abstract] Cancer cells and cancer associated stromal cells co-evolve secrete extracelluar vesicles to the surrounding regions and regulate several processes involved in cancer metastasis. miRNAs have been known to be mediators of cancer progression and metastasis. miRNAs consist of short noncoding RNA. miRNAs are stable in extracellular fluids such as serum, ...
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Scratch Wound Healing Assay

Author: Yanling Chen
Yanling ChenAffiliation: Department of Immunology, The Scripps Research Institute, La Jolla, USA
For correspondence: ylchen@scripps.edu
Bio-protocol author page: a27
date: 3/5/2012, 53944 views, 6 Q&A
DOI: https://doi.org/10.21769/BioProtoc.100.

[Abstract] The scratch wound healing assay has been widely adapted and modified to study the effects of a variety of experimental conditions, for instance, gene knockdown or chemical exposure, on mammalian cell migration and proliferation. In a typical scratch wound healing assay, a “wound gap” in a cell monolayer ...

Transwell Cell Migration Assay Using Human Breast Epithelial Cancer Cell

Author: Yanling Chen
Yanling ChenAffiliation: Department of Immunology, The Scripps Research Institute, La Jolla, USA
For correspondence: ylchen@scripps.edu
Bio-protocol author page: a27
date: 2/20/2012, 42738 views, 7 Q&A
DOI: https://doi.org/10.21769/BioProtoc.99.

[Abstract] Transwell migration assays have been widely used for studying the motility of different types of cells including metastatic cancer cells. The assay is also useful in screens for compounds that act as chemoattractants or inhibitors of chemotaxis for cells. The assay employs a permeable layer of support, ...

[Bio101] Cell Proliferation Assay by Flow Cytometry (BrdU and PI Staining)

Author: Hui Zhu
Hui ZhuAffiliation: Department of Genetics, Stanford University, Stanford, USA
For correspondence: huizhu@stanford.edu
Bio-protocol author page: a32
date: 4/5/2012, 38499 views, 2 Q&A
DOI: https://doi.org/10.21769/BioProtoc.198.

[Abstract] Cell Proliferation assays include an important set of fluorescence-based tests that can monitor cell health and cell division by evaluating DNA synthesis through thymidine incorporation. Bromodeoxyuridine (5-bromo-2'-deoxyuridine, BrdU) is a synthetic nucleoside that is an analogue of thymidine. BrdU ...

Clonogenic Assay

Author: Xiaodong Yang
Xiaodong YangAffiliation: Department of Neurology, University of California, San Francisco, USA
For correspondence: yangxiaodong1@yahoo.com
Bio-protocol author page: a43
date: 5/20/2012, 36971 views, 5 Q&A
DOI: https://doi.org/10.21769/BioProtoc.187.

[Abstract] Clonogenic assays serve as a useful tool to test whether a given cancer therapy can reduce the clonogenic survival of tumor cells. A colony is defined as a cluster of at least 50 cells that can often only be determined microscopically. A clonogenic assay is the method of choice to determine cell reproductive ...

In vitro Human Umbilical Vein Endothelial Cells (HUVEC) Tube-formation Assay

Authors: Josephine MY Ko
Josephine MY KoAffiliation: Clinical Oncology Department, The University of Hong Kong, Hong Kong , Hong Kong SAR
Bio-protocol author page: a100
 and Maria Li Lung
Maria Li LungAffiliation: Clinical Oncology Department, The Univerisity of Hong Kong, Hong Kong, Hong Kong SAR
For correspondence: mlilung@hku.hk
Bio-protocol author page: a101
date: 9/20/2012, 31960 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.260.

[Abstract] Angiogenesis is involved not only in pathological conditions including cancer biology and non-neoplastic diseases, but also many biological processes including reproduction, development and repair. During angiogenesis, endothelial cells (ECs) undergo activation after binding of angiogenic factors to ...

Soft–Agar colony Formation Assay

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: 7/5/2012, 30860 views, 6 Q&A
DOI: https://doi.org/10.21769/BioProtoc.220.

[Abstract] Any anchorage–independent growth of tumor cells is estimated by a soft–agar colony formation assay. This protocol provides a general workflow for establishing a soft-agar colony formation assay....

[Bio101] Cell Adhesion Assay

Author: Yanling Chen
Yanling ChenAffiliation: Department of Immunology, The Scripps Research Institute, La Jolla, USA
For correspondence: ylchen@scripps.edu
Bio-protocol author page: a27
date: 3/5/2012, 27733 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.98.

[Abstract] Cell adhesion, the binding of a cell to the extracellular matrix (ECM), other cells, or a specific surface, is essential for the growth and survival of the cell and also its communication with other cells. The process of cell adhesion involves a range of biological events such as three-dimensional re-organization ...

[Bio101] Subcutaneous Injection of Tumor Cells

Author: Jason Reuter date: 12/20/2011, 24323 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 vitro Tumorsphere Formation Assays

Authors: Sara Johnson
Sara JohnsonAffiliation: Biological Sciences Department, University of South Carolina, Columbia, USA
Bio-protocol author page: a224
Hexin Chen
Hexin ChenAffiliation: Biological Sciences Department, University of South Carolina, Columbia, USA
For correspondence: hchen@biol.sc.edu
Bio-protocol author page: a225
 and Pang-Kuo Lo
Pang-Kuo LoAffiliation: Biological Sciences Department, University of South Carolina, Columbia, USA
Bio-protocol author page: a226
date: 2/5/2013, 23813 views, 3 Q&A
DOI: https://doi.org/10.21769/BioProtoc.325.

[Abstract] A tumorsphere is a solid, spherical formation developed from the proliferation of one cancer stem/progenitor cell. These tumorspheres (Figure 1a) are easily distinguishable from single or aggregated cells (Figure 1b) as the cells appear to become fused together and individual cells cannot be identified. ...

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, 23204 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. ...
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