Accedi

University of Texas Southwestern Medical Center

39 ARTICLES PUBLISHED IN JoVE

image

Biology

Preparation of Developing and Adult Drosophila Brains and Retinae for Live Imaging
W. Ryan Williamson 1, P. Robin Hiesinger 1
1Department of Physiology and Green Center for Systems Biology, University of Texas Southwestern Medical Center

This protocol describes three Drosophila preparations: 1) adult brain dissection, 2) adult retina dissection and 3) developing eye disc- brain complexes dissection. Emphasis is laid on special preparation techniques and conditions for live imaging, although all preparations can be used for fixed tissue immunohistochemistry.

image

Biology

Method for the Isolation of Francisella tularensis Outer Membranes
Jason F. Huntley 1, Gregory T. Robertson 1, Michael V. Norgard 1
1Department of Microbiology, University of Texas Southwestern Medical Center

A protocol for separating inner and outer membranes from Francisella tularensis by spheroplasting, osmotic lysis, and sucrose density gradient ultracentrifugation.

image

Neuroscience

The Vermicelli and Capellini Handling Tests: Simple quantitative measures of dexterous forepaw function in rats and mice
Kelly A. Tennant 1, Aaron L. Asay 2, Rachel P. Allred 3, Angela R. Ozburn 4, Jeffrey A. Kleim 5, Theresa A. Jones 1,2
1Institute for Neuroscience, University of Texas at Austin, 2Department of Psychology, University of Texas at Austin, 3Department of Neurology, University of Florida, 4Department of Psychiatry, University of Texas Southwestern Medical Center, 5Department of Neuroscience, McKnight Brain Institute, University of Florida

The Vermicelli and Capellini Handling Tests of forepaw dexterity take advantage of the natural inclination of rodents to manipulate food items using skillful forepaw and digit movements. Animals are videotaped while handling short strands of uncooked dry pasta. Slow motion video playback allows for the quantification of forepaw adjustments.

image

Neuroscience

Preparation of Acute Hippocampal Slices from Rats and Transgenic Mice for the Study of Synaptic Alterations during Aging and Amyloid Pathology
Diana M. Mathis 1, Jennifer L. Furman 2, Christopher M. Norris 2,3
1Graduate Center for Gerontology, University of Kentucky College of Public Health, 2Department of Molecular and Biomedical Pharmacology, University of Kentucky College of Medicine, 3Sanders-Brown Center on Aging, University of Kentucky College of Medicine

This article outlines procedures for preparing hippocampal slices from rats and transgenic mice for the study of synaptic alterations associated with brain aging and age-related neurodegenerative diseases, such as Alzheimer’s disease.

image

Medicine

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model
Debabrata Saha 1, Henry Dunn 2, Heling Zhou 2, Hiroshi Harada 3, Masahiro Hiraoka 3, Ralph P. Mason 2, Dawen Zhao 2
1Department of Radiation Oncology, University of Texas Southwestern Medical Center , 2Department of Radiology, University of Texas Southwestern Medical Center , 3Department of Radiation Oncology, Kyoto University Graduate School of Medicine

Bioluminescence imaging of hypoxia inducible factor-1α activity is applied to monitor intracranial tumor hypoxia development in a breast cancer brain metastasis mouse model.

image

Biology

Recombineering Homologous Recombination Constructs in Drosophila
Arnaldo Carreira-Rosario 1, Shane Scoggin 1, Nevine A. Shalaby 1, Nathan David Williams 1, P. Robin Hiesinger 2,3, Michael Buszczak 1
1Department of Molecular Biology, University of Texas Southwestern Medical Center at Dallas, 2Department of Physiology, University of Texas Southwestern Medical Center at Dallas, 3Green Center for Systems Biology, University of Texas Southwestern Medical Center at Dallas

Homologous recombination techniques greatly advance Drosophila genetics by enabling the creation of molecularly precise mutations. The recent adoption of recombineering allows one to manipulate large pieces of DNA and transform them into Drosophila6. The methods presented here combine these techniques to rapidly generate large homologous recombination vectors.

image

Medicine

Ultrasound Imaging-guided Intracardiac Injection to Develop a Mouse Model of Breast Cancer Brain Metastases Followed by Longitudinal MRI
Heling Zhou 1, Dawen Zhao 1
1Radiology, University of Texas Southwestern Medical Center

A breast cancer brain metastasis mouse model is established with ultrasound imaging-guided intracardiac injection of MDA-MB231/Br-GFP cells. Development of multifocal intracranial metastases has been monitored longitudinally using high-resolution 9.4 T MRI.

image

Medicine

MRI Mapping of Cerebrovascular Reactivity via Gas Inhalation Challenges
Hanzhang Lu 1, Peiying Liu 1, Uma Yezhuvath 1, Yamei Cheng 1, Olga Marshall 2, Yulin Ge 2
1Advanced Imaging Research Center, University of Texas Southwestern Medical Center, 2Center for Biomedical Imaging, Department of Radiology, New York University School of Medicine

Non-invasive imaging of the brain vasculature’s ability to dilate or constrict may allow a better understanding of cerebrovascular pathophysiology in various neurological diseases. The present report describes a reproducible and patient-comfortable protocol to perform vascular reactivity imaging in humans using magnetic resonance imaging (MRI).

image

Medicine

Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice
Katherine Poinsatte 1, Uma Maheswari Selvaraj 1, Sterling B. Ortega 1, Erik J. Plautz 1, Xiangmei Kong 1, Jeffrey M. Gidday 2, Ann M. Stowe 1
1Department of Neurology and Neurotherapeutics, University of Texas Southwestern Medical Center, 2Department of Neurological Surgery, Washington University School of Medicine

This protocol describes repetitive hypoxic preconditioning, or brief exposures to systemic hypoxia that reduce infarct volumes and blood-brain barrier disruption following transient middle cerebral artery occlusion in mice. It also details dual quantification of infarct volume and blood-brain barrier disruption after stroke to assess the efficacy of neurovascular protection.

image

Biology

Use of the Ramsay Assay to Measure Fluid Secretion and Ion Flux Rates in the Drosophila melanogaster Malpighian Tubule
Jeffrey N. Schellinger 1, Aylin R. Rodan 1
1Department of Internal Medicine, University of Texas Southwestern Medical Center

This protocol describes the use of the Ramsay assay to measure fluid secretion rates from isolated Malpighian (renal) tubules from Drosophila melanogaster. In addition, the use of ion-specific electrodes to measure sodium and potassium concentrations in the secreted fluid, allowing calculation of transepithelial ion flux, is described.

image

Bioengineering

Sensitive Detection of Proteopathic Seeding Activity with FRET Flow Cytometry
Jennifer L. Furman *1, Brandon B. Holmes *1, Marc I. Diamond 1
1Center for Alzheimer's and Neurodegenerative Diseases, University of Texas Southwestern Medical Center

Cell-to-cell transfer of protein aggregates, or proteopathic seeds, may underlie the progression of pathology in neurodegenerative diseases. Here, a novel FRET flow cytometry assay is described that enables specific and sensitive detection of seeding activity from recombinant or biological samples.

image

Engineering

Use of a Multi-compartment Dynamic Single Enzyme Phantom for Studies of Hyperpolarized Magnetic Resonance Agents
Christopher M. Walker 1, Matthew Merritt 2, Jian-Xiong Wang 2, James A. Bankson 1
1Imaging Physics, University of Texas M.D. Anderson Cancer Center, 2Advanced Imaging Research Center, University of Texas Southwestern Medical Center

A multi-compartment dynamic phantom is used to simulate some biology of interest for metabolic studies using hyperpolarized magnet resonance agents.

image

Neuroscience

Whole-cell Patch-clamp Recordings in Brain Slices
Amir Segev 1, Francisco Garcia-Oscos 1, Saïd Kourrich 1
1Department of Psychiatry, University of Texas Southwestern Medical Center

This protocol describes basic procedural steps for performing whole-cell patch-clamp recordings. This technique allows the study of the electrical behavior of neurons, and when performed in brain slices, allows the assessment of various neuronal functions from neurons that are still integrated in relatively well preserved brain circuits.

image

Biology

A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing
Michael Allen Stiffler 1, Subu K Subramanian 1, Victor H Salinas 1, Rama Ranganathan 1
1Green Center for Systems Biology, University of Texas Southwestern Medical Center

We present a protocol for the functional assessment of comprehensive single-site saturation mutagenesis libraries of proteins utilizing high-throughput sequencing. Importantly, this approach uses orthogonal primer pairs to multiplex library construction and sequencing. Representative results using TEM-1 β-lactamase selected at a clinically relevant dosage of ampicillin are provided.

image

Developmental Biology

Using Primary Neurosphere Cultures to Study Primary Cilia
Issei S. Shimada 1, Hemant Badgandi 1, Bandarigoda N. Somatilaka 1, Saikat Mukhopadhyay 1
1Department of Cell Biology, University of Texas Southwestern Medical Center

The primary cilium is fundamentally important in neural progenitor cell proliferation, neuronal differentiation, and adult neuronal function. Here, we describe a method to study ciliogenesis and the trafficking of signaling proteins to cilia in neural stem/progenitor cells and differentiated neurons using primary neurosphere cultures.

image

JoVE Core

Assaying Protein Kinase Activity with Radiolabeled ATP
Aroon S. Karra 1, Steve Stippec 1, Melanie H. Cobb 1
1Department of Pharmacology, University of Texas Southwestern Medical Center

Protein kinases are highly evolved signaling enzymes and scaffolds that are critical for inter- and intracellular signal transduction. We present a protocol for measuring kinase activity through the use of radiolabeled adenosine triphosphate ([γ-32P] ATP), a reliable method to aid in elucidation of cellular signaling regulation.

image

Medicine

Tubal Cytology of the Fallopian Tube as a Promising Tool for Ovarian Cancer Early Detection
Hao Chen 1, Robert Klein 1, Stacy Arnold 1, Yiying Wang 2, Setsuko Chambers 3,4, Wenxin Zheng 5,6
1Department of Pathology, University of Arizona College of Medicine, 2Department of Obstetrics and Gynecology, Henan Provincial People's Hospital, 3Department of Obstetrics and Gynecology, University of Arizona College of Medicine, 4University of Arizona Cancer Center, 5Department of Pathology, University of Texas Southwestern Medical Center, 6Department of Obstetrics and Gynecology, University of Texas Southwestern Medical Center

We explored a tubal cytologic method by sampling the fallopian tube directly post-surgical excision as a tool of ovarian cancer early detection. Here, we present a protocol to collect fallopian tube cells from freshly received surgical specimens.

image

Immunology and Infection

Arbovirus Infections As Screening Tools for the Identification of Viral Immunomodulators and Host Antiviral Factors
Emily A. Rex 1, Dahee Seo 1, Don B. Gammon 1
1Department of Microbiology, University of Texas Southwestern Medical Center

Here, we present the protocols to identify 1) virus-encoded immunomodulators that promote arbovirus replication and 2) eukaryotic host factors that restrict arbovirus replication. These fluorescence- and luminescence-based methods allow researchers to rapidly obtain quantitative readouts of arbovirus replication in simplistic assays with low signal-to-noise ratios.

image

Biology

Measuring Relative Insulin Secretion using a Co-Secreted Luciferase Surrogate
Michael Kalwat 1, Melanie H. Cobb 1
1Department of Pharmacology, University of Texas Southwestern Medical Center

This protocol describes how to perform rapid low-cost luciferase assays at medium-throughput using an insulin-linked Gaussia luciferase as a proxy for insulin secretion from beta cells. The assay can be performed with most luminescence plate readers and multichannel pipettes.

image

Neuroscience

Hybrid Microdrive System with Recoverable Opto-Silicon Probe and Tetrode for Dual-Site High Density Recording in Freely Moving Mice
Hisayuki Osanai 1, Takashi Kitamura 1,2, Jun Yamamoto 1
1Department of Psychiatry, University of Texas Southwestern Medical Center, 2Department of Neuroscience, University of Texas Southwestern Medical Center

This protocol describes the construction of a hybrid microdrive array that allows implantation of nine independently adjustable tetrodes and one adjustable opto-silicon probe in two brain regions in freely moving mice. Also demonstrated is a method for safely recovering and reusing the opto-silicon probe for multiple purposes.

image

Biology

Detection of Tissue-resident Bacteria in Bladder Biopsies by 16S rRNA Fluorescence In Situ Hybridization
Michael L. Neugent *1, Jashkaran Gadhvi *1, Kelli L. Palmer 1, Philippe E. Zimmern 2, Nicole J. De Nisco 1
1Department of Biological Sciences, University of Texas at Dallas, 2Department of Urology, University of Texas Southwestern Medical Center

This protocol is for the unbiased detection of tissue-associated bacteria in patient biopsies by 16S rRNA in situ hybridization and confocal microscopy.

image

Genetics

Transcriptome-Wide Profiling of Protein-RNA Interactions by Cross-Linking and Immunoprecipitation Mediated by FLAG-Biotin Tandem Purification
Xianju Bi 1,2, Xuechun Zhang 1, Xiaohua Shen 1
1Tsinghua-Peking Center for Life Sciences, School of Medicine and School of Life Sciences, Tsinghua University, 2Department of Molecular Biology, University of Texas Southwestern Medical Center

Here we present a modified CLIP-seq protocol called FbioCLIP-seq with FLAG-biotin tandem purification to determine the RNA targets of RNA-binding proteins (RBPs) in mammalian cells.

image

Immunology and Infection

Isolation of Adipogenic and Fibro-Inflammatory Stromal Cell Subpopulations from Murine Intra-Abdominal Adipose Depots
Julia Peics *1, Lavanya Vishvanath *2, Qianbin Zhang 2, Bo Shan 2, Thomas Å. Pedersen 1, Rana K. Gupta 2
1Diabetes Pharmacology, Novo Nordisk A/S, 2Touchstone Diabetes Center, Department of Internal Medicine, University of Texas Southwestern Medical Center

This protocol describes the technical approach to isolate adipogenic and fibro-inflammatory stromal cell subpopulations from murine intra-abdominal white adipose tissue (WAT) depots by fluorescence-activated cell sorting or immunomagnetic bead separation.

image

Biology

Cryogenic Sample Loading into a Magic Angle Spinning Nuclear Magnetic Resonance Spectrometer that Preserves Cellular Viability
Rupam Ghosh *1, Jaka Kragelj *1, Yiling Xiao *1, Kendra K. Frederick 1,2
1Department of Biophysics, University of Texas Southwestern Medical Center, 2Center for Alzheimer's and Neurodegenerative Disease and Center for Systems Biology, University of Texas Southwestern Medical Center

Presented here is a protocol for cryogenic transfer of frozen samples into the dynamic nuclear polarization (DNP) magic angle spinning (MAS) nuclear magnetic resonance (NMR) probe. The protocol includes directions for rotor storage prior to the experiment and directions for viability measurements before and after the experiment.

image

Biochemistry

Analysis of Neutral Lipid Synthesis in Saccharomyces cerevisiae by Metabolic Labeling and Thin Layer Chromatography
Sean Rogers 1, W. Mike Henne 1
1Department of Cell Biology, University of Texas Southwestern Medical Center

Here, a protocol is presented for the metabolic labeling of yeast with 14C-acetic acid, which is coupled with thin layer chromatography for the separation of neutral lipids.

image

Genetics

Hybrid De Novo Genome Assembly for the Generation of Complete Genomes of Urinary Bacteria using Short- and Long-read Sequencing Technologies
Belle M. Sharon 1, Neha V. Hulyalkar 1, Vivian H. Nguyen 1, Philippe E. Zimmern 2, Kelli L. Palmer 1, Nicole J. De Nisco 1
1Department of Biological Sciences, University of Texas at Dallas, 2Department of Urology, University of Texas Southwestern Medical Center

This protocol details a comprehensive approach for the culturing, sequencing, and de novo hybrid genome assembly of urinary bacteria. It provides a reproducible procedure for the generation of complete, circular genome sequences useful in studying both chromosomal and extrachromosomal genetic elements contributing to urinary colonization, pathogenesis, and antimicrobial resistance dissemination.

image

Cancer Research

Analytical Determination of Mitochondrial Function of Excised Solid Tumor Homogenates
Elizabeth R. M. Zunica 1,2,3, Christopher L. Axelrod 1, L. Anne Gilmore 2,4, John P. Kirwan 1,3
1Integrated Physiology and Molecular Medicine Laboratory, Pennington Biomedical Research Center, 2Clinical Oncology and Metabolism, Pennington Biomedical Research Center, 3Department of Nutrition, Case Western Reserve University, 4Department of Clinical Nutrition, University of Texas Southwestern Medical Center

We developed a practical protocol and analytical approach to evaluate mitochondrial oxidative phosphorylation and electron transfer capacity in fresh tumor homogenates. This protocol can be easily adapted to survey various mitochondrial functions that contribute to cancer initiation, progression, and treatment response.

image

Medicine

Evaluation of Amino Acid Consumption in Cultured Bone Cells and Isolated Bone Shafts
Leyao Shen 1, Courtney M. Karner 1,2
1Department of Internal Medicine, University of Texas Southwestern Medical Center, 2Charles and Jane Pak Center for Mineral Metabolism and Clinical Research, University of Texas Southwestern Medical Center

This protocol presents a radiolabeled amino acid uptake assay, which is useful for evaluating amino acid consumption either in primary cells or in isolated bones.

image

JoVE Journal

Monitoring Gut Acidification in the Adult Drosophila Intestine
Farhan Abu 1, Benjamin Ohlstein 1
1Children's Research Institute, University of Texas Southwestern Medical Center

Here, we present a standardized protocol for monitoring gut acidification in Drosophila melanogaster with optimal output. We first use this protocol for gut acidification monitoring in Drosophila melanogaster and then demonstrate its use in non-model Drosophila species.

image

Medicine

Reduced Procedure Time and Variability with Active Esophageal Cooling During Radiofrequency Ablation for Atrial Fibrillation
Christopher Joseph 1, Julie Cooper 2, Robert W. Turer 3, Samuel A. McDonald 3, Erik Benjamin Kulstad 1, James Daniels 4
1Department of Emergency Medicine, University of Texas Southwestern Medical Center, 2Department of Internal Medicine, University of Texas Southwestern Medical Center, 3Department of Emergency Medicine and Clinical Informatics Center, University of Texas Southwestern Medical Center, 4Department of Electrophysiology, University of Texas Southwestern Medical Center

This study utilized advanced informatics techniques to compare procedure duration in patients undergoing radiofrequency atrial ablation treated with active esophageal cooling to those treated with traditional luminal esophageal temperature monitoring. Contextual inquiry, workflow analysis, and data mapping were utilized. The findings demonstrated reduced procedure time and variability with active cooling.

image

Medicine

Partial Sciatic Nerve Ligation: A Mouse Model of Chronic Neuropathic Pain to Study the Antinociceptive Effect of Novel Therapies
Hannah E. Korah 1, Kevin Cheng 1, Stephanie M. Washington 2, Matthew E. Flowers 1, Harrison J. Stratton 1, Amol Patwardhan 5, Mohab M. Ibrahim *1,2,3,4, Laurent F. Martin *1,2
1Department of Pharmacology, College of Medicine, The University of Arizona, Tucson, 2Department of Anesthesiology, College of Medicine, The University of Arizona, Tucson, 3Neuroscience Graduate Interdisciplinary Program, College of Medicine, The University of Arizona, Tucson, 4Comprehensive Pain and Addiction Center, The University of Arizona, Tucson, 5Department of Anesthesiology and Pain Management, University of Texas Southwestern Medical Center

Partial sciatic nerve ligation induces long-lasting chronic neuropathic pain, characterized by exaggerated responses to thermal and mechanical stimuli. This mouse model of neuropathic pain is commonly used to study innovative therapies for pain management. This article describes in detail the surgical procedure to improve standardization and reproducibility.

image

Cancer Research

Generating and Imaging Mouse and Human Epithelial Organoids from Normal and Tumor Mammary Tissue Without Passaging
Serena L. Cornelius 1,2, Megan M. Colonnetta 2,3,4,5, Katherine E. Lake 1,2, Clayton A. Smith 1,2, Yu-An Zhang 1,2, Evanthia T. Roussos-Torres 6, Sangeetha M. Reddy 1,2, Elizabeth H. Chen 2,3,4,5, Isaac S. Chan 1,2,3,4
1Department of Internal Medicine, Division of Hematology and Oncology, University of Texas Southwestern Medical Center, 2Harold C. Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, 3Department of Molecular Biology, University of Texas Southwestern Medical Center, 4Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center, 5Department of Cell Biology, University of Texas Southwestern Medical Center, 6Division of Medical Oncology, Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California

This protocol discusses an approach for generating epithelial organoids from primary normal and tumor mammary tissue through differential centrifugation. Furthermore, instructions are included for three-dimensional culturing as well as immunofluorescent imaging of embedded organoids.

image

Methods For Studying Osteoenergetics And Metabolism
Elizabeth Rendina-Ruedy 1, Courtney M. Karner 2
1Department of Medicine, Clinical Pharmacology, Vanderbilt University Medical Center, 2Department of Internal Medicine, Charles and Jane Pak Center for Mineral Metabolism and Clinical Research, University of Texas Southwestern Medical Center

Methods For Studying Osteoenergetics And Metabolism

image

Biochemistry

An Acetyl-Click Chemistry Assay to Measure Histone Acetyltransferase 1 Acetylation
Shreenidhi Rajkumar 1, Danielle Dixon 1, Andrew M. Lipchik 2, Joshua J. Gruber 1
1Departments of Internal Medicine and the Green Center for Reproductive Biology Sciences, University of Texas Southwestern Medical Center, 2Department of Pharmaceutical Sciences, Eugene Applebaum College of Pharmacy and Health Sciences, Wayne State University

Quick and accurate chemical assays to screen for specific inhibitors are an important tool in the drug development arsenal. Here, we present a scalable acetyl-click chemistry assay to measure the inhibition of HAT1 acetylation activity.

image

Medicine

Determination of Continuity Index Values in Atrial Fibrillation Ablation with Proactive Esophageal Cooling
Catherine Lazarus 1, Jacob Sherman 2, Natalie Putzel 3, Tiffany Sharkoski 4, William Zagrodzky 4, Erik Kulstad 4,5, Alex Ro 6, Jose Nazari 6, Westby Fisher 6, Mark Metzl 6
1Northwestern University, 2Washington University in St. Louis, 3University of Southern California, 4Attune Medical, 5University of Texas Southwestern Medical Center, 6NorthShore University Hospital

This protocol describes the methods used to determine the continuity index in patients undergoing pulmonary vein isolation procedures using radiofrequency ablation and demonstrates the differences in continuity index between ablation procedures using proactive esophageal cooling as compared to procedures using traditional luminal esophageal temperature monitoring.

image

Neuroscience

Monitoring Blood-Brain Barrier Opening in Rats with a Preclinical Focused Ultrasound System
Kisoo Kim 1,2, Marco Gallus 3, Tianrun Xiao 1, Akshay S. Parchure 4, Bhavya R. Shah 4, Hideho Okada 3, Chris Diederich 5, Eugene Ozhinsky *1, Kazim Narsinh *1
1Department of Radiology & Biomedical Imaging, University of California, San Francisco, 2Department of Biomedical Engineering, Kyung Hee University, 3Department of Neurological Surgery, University of California, San Francisco, 4University of Texas Southwestern Medical Center, 5Department of Radiation Oncology, University of California, San Francisco

This study describes a combined magnetic resonance imaging (MRI) and low-intensity pulsed focused ultrasound (FUS) protocol, utilizing living rats with jugular vein catheterization to monitor blood-brain barrier (BBB) opening.

image

Medicine

Robotic-assisted Bronchoscopy Combined with Multimodal Imaging for Targeted Lung Cryobiopsies
David Pham 1, Kim Styrvoky 1
1Division of Pulmonary and Critical Care Medicine, University of Texas Southwestern Medical Center

This article aims to describe a stepwise approach to performing robotic-assisted bronchoscopy combined with fluoroscopy, radial endobronchial ultrasound, and cone beam computed tomography to obtain targeted transbronchial lung cryobiopsies. 

image

Cancer Research

Robust Detection of Gene Amplification in Formalin-Fixed Paraffin-Embedded Samples by Fluorescence In Situ Hybridization
Collin Gilbreath 1, Yan Peng 2, Sihan Wu 1
1Children's Medical Center Research Institute, University of Texas Southwestern Medical Center, 2Department of Pathology, University of Texas Southwestern Medical Center

This protocol provides a reproducible method to visualize gene amplification in formalin-fixed paraffin-embedded (FFPE) tissue specimens.

image

Biochemistry

Utilizing Thermal Shift Assay to Probe Substrate Binding to Selenoprotein O
Abner Gonzalez 1, Anju Sreelatha 1,2
1Department of Physiology, University of Texas Southwestern Medical Center, 2Charles and Jane Pak Center for Mineral Metabolism and Clinical Research, University of Texas Southwestern Medical Center

Here, we present the thermal shift assay, a high-throughput, fluorescence-based technique used to investigate the binding of small molecules to proteins of interest.

JoVE Logo

Riservatezza

Condizioni di utilizzo

Politiche

Ricerca

Didattica

CHI SIAMO

Copyright © 2024 MyJoVE Corporation. Tutti i diritti riservati