Cilia. Part A /

This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers cilia and includes chapters on such topics as electron microscopy of IFT in cilia and flagella, radial spoke isolation and assays, and biome...

Πλήρης περιγραφή

Λεπτομέρειες βιβλιογραφικής εγγραφής
Άλλοι συγγραφείς: Marshall, Wallace F.
Μορφή: Ηλ. βιβλίο
Γλώσσα:English
Έκδοση: San Diego : Elsevier Science, 2013.
Σειρά:Methods in enzymology ; v. 524.
Θέματα:
Διαθέσιμο Online:Full Text via HEAL-Link
LEADER 07018nam a2200613 4500
001 ocn831118382
003 OCoLC
005 20180501122008.0
006 m o d
007 cr mn|||||||||
008 130323s2013 cauab oab 001 0 eng d
040 |a EBLCP  |b eng  |e pn  |c EBLCP  |d MBB  |d BUF  |d N$T  |d MKC  |d OPELS  |d OCLCF  |d UKDOC  |d VVL  |d KUK  |d OCLCQ  |d YDXCP  |d DEBSZ  |d E7B  |d OCLCQ  |d FEM  |d OCLCQ  |d TFH  |d OCLCO  |d NJR  |d OCLCO  |d OCLCA  |d GrThAP 
019 |a 836202791  |a 839295030  |a 839296147  |a 841170349  |a 968094267  |a 969011813 
020 |a 9780123984869  |q (electronic bk.) 
020 |a 0123984866  |q (electronic bk.) 
020 |a 9780123979452  |q (print) 
020 |a 0123979455  |q (print) 
035 |a (OCoLC)831118382  |z (OCoLC)836202791  |z (OCoLC)839295030  |z (OCoLC)839296147  |z (OCoLC)841170349  |z (OCoLC)968094267  |z (OCoLC)969011813 
050 4 |a QH647 .C384 2013 
060 4 |a QU135  |b .M49 v.524 
072 7 |a SCI  |x 017000  |2 bisacsh 
082 0 4 |a 571.67 
049 |a TEFA 
245 0 0 |a Cilia.  |n Part A /  |c edited by Wallace F. Marshall. 
260 |a San Diego :  |b Elsevier Science,  |c 2013. 
300 |a 1 online resource (469 pages) :  |b illustrations (some color). 
336 |a text  |b txt  |2 rdacontent 
337 |a computer  |b c  |2 rdamedia 
338 |a online resource  |b cr  |2 rdacarrier 
347 |a text file  |2 rda 
490 1 |a Methods in enzymology ;  |v v. 524.  |x 0076-6879 
588 0 |a Print version record. 
504 |a Includes bibliographical references and index. 
520 |a This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers cilia and includes chapters on such topics as electron microscopy of IFT in cilia and flagella, radial spoke isolation and assays, and biomechanical measurements of kinocilium. Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers ciliaContains chapters on such topics as electron microscopy of IFT in cilia and flagella, radial spoke isolation. 
505 0 |a Isolation of Intraflagellar Transport Particle Proteins from Chlamydomonas reinhardtii -- Molecular Tools for Studying the Radial Spoke -- Methods for Analysis of Calcium/Calmodulin Signaling in Cilia and Flagella -- Analysis of Flagellar Protein Ubiquitination -- Analysis of Ciliary Import -- Analysis of Signaling Pathways Controlling Flagellar Movements in Mammalian Spermatozoa -- Analysis of Microtubule Plus-End-Tracking Proteins in Cilia -- Biochemical and Physiological Analysis of Axonemal Dyneins -- Measuring the Regulation of Dynein Activity During Flagellar Motility -- Analysis of Interactions Between Intraflagellar Transport Proteins -- Analysis of Ciliary Membrane Protein Dynamics Using SNAP Technology -- Engineering Oscillating Microtubule Bundles -- Purification of Exosome-Like Vesicles from Urine -- Electron Microscopy of Flagella, Primary Cilia, and Intraflagellar Transport in Flat-Embedded Cells -- In vivo Imaging of IFT in Chlamydomonas Flagella -- Measuring Rates of Intraflagellar Transport Along Caenorhabditis elegans Sensory Cilia Using Fluorescence Microscopy -- 3D Structural Analysis of Flagella/Cilia by Cryo-Electron Tomography -- Electron Tomography of IFT Particles -- Reconstitution of Flagellar Sliding. 
505 0 |a Front Cover; Cilia, Part A; Copyright; Contents; Contributors; Preface; Methods in Enzymology; Chapter One: Isolation of Intraflagellar Transport Particle Proteins from Chlamydomonas reinhardtii; 1. Introduction; 2. Media; 2.1. Tips on making media; 2.2. Media choices; 3. Isolating Flagella; 3.1. Recipe of stock solutions (the chemicals are from Sigma); 3.2. Preparing the culture; 3.3. Isolating flagella; 4. Preparing Soluble Flagellar Proteins; 5. Isolating IFT Particle Proteins by Fractioning Soluble Flagellar Proteins Through Sucrose Density Gradient. 
505 8 |a 5.1. Prepare low- and high-density sucrose solutions5.2. Making the gradient; 5.3. Gradient centrifugation; 5.4. Collecting the gradient fractions; 5.5. Analyzing the fractions by gel electrophoresis; 6. Isolating IFT Particle Proteins by Immunoprecipitation from Soluble Flagellar Proteins; 7. Conclusions; Acknowledgments; References; Chapter Two: Molecular Tools for Studying the Radial Spoke; 1. Introduction; 2. Recovery of Genomic DNA; 2.1. PCR genomic DNA; 2.2. Random subcloning of BAC DNA; 3. Slot Lysis Assay for Large-Scale Screenings; Solutions; 4. DNA Alterations; 5. Tagging. 
505 8 |a 5.1. Add a tag DNA to a gene5.2. Fuse a short DNA fragment to a tag DNA; 6. Ni-NTA Affinity Purification of the Radial Spoke Complex; 7. Flagella Miniprep; Solutions; 8. Summary; Acknowledgments; References; Chapter Three: Methods for Analysis of Calcium/Calmodulin Signaling in Cilia and Flagella; 1. Introduction; 2. Calcium Entry into the Intraflagellar Space; 3. Calcium-Binding Proteins; 3.1. Identifying calcium sensors by molecular and biochemical methods; 3.1.1. Electrophoretic mobility shift and phenyl-sepharose binding; 3.1.2. Radioisotope 45-Ca2+ 
505 8 |a 3.2. Identifying calmodulin-binding proteins3.2.1. Affinity purification using calmodulin-coupled sepharose; 3.2.2. Calmodulin gel overlay assay; 3.2.2.1. Materials; 3.2.2.2. Method; 3.2.3. Calmodulin immunoprecipitation; 3.2.3.1. Materials; 3.2.3.2. Method; 4. Calcium-Induced Modulation of Motility; 4.1. Inducing and imaging phototaxis and photoshock in C. reinhardtii; 4.2. Analyzing photo-accumulation; 4.3. Reactivation studies using cell models or isolated axonemes; 4.4. Microtubule sliding assay; 4.4.1. Materials; 4.4.2. Method; 5. Summary; References. 
505 8 |a Chapter Four: Analysis of Flagellar Protein Ubiquitination1. Introduction; 2. Summary of Putative Ubiquitin, Ubiquitin-Like Proteins, E1, E2, and E3 in Flagella/Cilia; 3. Determination of the Flagellar Localization of Candidate Proteins with Fluorescence Proteins; 4. Analysis of the Free Ubiquitin in Flagella; 5. In Vitro Ubiquitination System of Flagella; Reagents; Procedures; 6. Analysis of Flagellar Ubiquitination During Flagellar Shortening; Reagents; Procedures; Procedures; 7. Analysis of Flagellar Ubiquitination During Mating; Procedures; 8. Summary; Acknowledgment; References. 
650 0 |a Cilia and ciliary motion. 
650 0 |a Flagella (Microbiology) 
650 0 |a Cells  |x Motility. 
650 7 |a SCIENCE  |x Life Sciences  |x Cell Biology.  |2 bisacsh 
650 7 |a Cells  |x Motility.  |2 fast  |0 (OCoLC)fst00850268 
650 7 |a Cilia and ciliary motion.  |2 fast  |0 (OCoLC)fst00861419 
650 7 |a Flagella (Microbiology)  |2 fast  |0 (OCoLC)fst00926920 
650 2 |a Cilia  |x physiology. 
655 4 |a Electronic books. 
655 7 |a Electronic books.  |2 lcgft 
700 1 |a Marshall, Wallace F. 
776 0 8 |i Print version:  |a Marshall, Wallace F.  |t Cilia, Part A.  |d San Diego : Elsevier Science, ©2013  |z 9780123979452 
830 0 |a Methods in enzymology ;  |v v. 524.  |x 0076-6879 
856 4 0 |u https://www.sciencedirect.com/science/bookseries/00766879/524  |z Full Text via HEAL-Link