|
- 积分
- 1164
- 威望
- 1164
- 包包
- 245
|
本帖最后由 细胞海洋 于 2013-1-24 14:01 编辑
7 X: L6 U2 L0 [1 Z) l
, i/ I6 S$ z7 c- O! f0 TCurrent Protocols in Cell Biology 2010年完整版 5483页4 h5 n( `( I/ D% e S, J
( L+ A4 X0 Y9 b+ A a( POnline ISBN: 97804711430311 f6 ?( F$ \) T8 U
DOI: 10.1002/0471143030% C0 ]. P. u, l* \8 d1 t. j; I
/ m3 c6 m5 {6 j1 UTable of Contents
; V7 k0 l% E; |9 G& Y- u7 H: c1. Preface
1 s6 _: `' J; u& j+ [2. Foreword
9 f5 h& m; M0 A9 `2 X3. Chapter 1 Cell Culture
3 N- }3 U& P8 F4 b! u) t1. Introduction2 Z; v" n, h1 K9 _
2. Unit 1.1 Basic Techniques in Mammalian Cell Tissue Culture) L, U8 k2 \, }5 Q8 h
3. Unit 1.2 Media for Culture of Mammalian Cells
3 z1 x: H$ k! p4. Unit 1.3 Aseptic Technique for Cell Culture
) j9 K) U2 S& i1 e ~5. Unit 1.4 Sterilization and Filtration
& [! W3 ~6 A9 [0 x; O v4 v& t6. Unit 1.5 Assessing and Controlling Microbial Contamination in Cell Cultures
* H2 V" i( m. _' w1 q7. Unit 1.6 Media and Culture of Yeast0 x' ]# G. G4 N7 R; d- G- L$ B
8. Unit 1.7 BY-2 Cells: Culture and Transformation for Live Cell Imaging
# c7 G+ g/ q# P" F; i. [# N; k+ ]4. Chapter 2 Preparation and Isolation of Cells# Z9 @% R+ B. T$ x& \8 |0 Z& A
1. Introduction
6 p- h. {3 a" P, {4 Z2. Unit 2.1 Establishment of Fibroblast Cultures/ P" Q7 t% h9 s g4 K6 w
3. Unit 2.2 Preparation and Culture of Human Lymphocytes+ h I% M; p& X8 N4 ^
4. Unit 2.3 Preparation of Endothelial Cells
0 V' l- C1 z8 W1 D! N5. Unit 2.4 Generation of Continuously Growing B Cell Lines by Epstein-Barr Virus Transformation
1 `4 O1 L4 N% C2 E9 c# o8 f6. Unit 2.5 Laser Capture Microdissection Y# T& {6 k! O! b' r/ ~0 M9 W+ T
7. Unit 2.6 Preparation of Human Epidermal Keratinocyte Cultures
6 }- m- U; _8 Y4 S" Y$ F8. Unit 2.7 Preparation and Coculture of Neurons and Glial Cells* }: k& H9 Q4 j
5. Chapter 3 Subcellular Fractionation and Isolation of Organelles
( ?" J. Q0 b: ~6 h8 I1. Introduction
* C a* h D) }. S" N4 {2. Introduction( ]- T4 Y& J8 b# a$ R
3. Unit 3.1 Overview of Cell Fractionation
2 A( `% f) X8 L1 q) x4. Unit 3.2 Isolation of Rat Hepatocyte Plasma Membrane Sheets and Plasma Membrane Domains
}, O3 ?2 p8 D4 H5. Unit 3.3 Isolation of Mitochondria from Tissues and Cells by Differential Centrifugation
* w' j |& Y' q" S' P8 N6. Unit 3.4 Purification of a Crude Mitochondrial Fraction by Density-Gradient Centrifugation7 I2 [+ H; s3 [- W2 j0 g
7. Unit 3.5 Isolation of Peroxisomes from Tissues and Cells by Differential and Density Gradient2 L2 I* q% N7 }/ h7 S
Centrifugation; c0 A n4 a7 Z5 B' _
8. Unit 3.6 Isolation of Lysosomes from Tissues and Cells by Differential and Density Gradient# c5 l3 h' e" @! o! s# Y
Centrifugation" G' H) o7 C/ s. B0 d/ a# C
9. Unit 3.7 Overview of Subcellular Fractionation Procedures for the Yeast Saccharomyces cerevisiae
l2 u/ g. s( r8 _10. Unit 3.8 Isolation of Subcellular Fractions from the Yeast Saccharomyces cerevisiae
4 s+ u! V) W o9 K6 ~11. Unit 3.9 Isolation of Golgi Membranes from Tissues and Cells by Differential and Density Gradient
2 X, N& `3 E7 j2 cCentrifugation$ H; i7 Z0 F5 n, B7 p$ q2 L
12. Unit 3.10 Isolation of Nuclei and Nuclear Membranes From Animal Tissues. A% ~3 _ c! g: h
13. Unit 3.11 Free-Flow Electrophoretic Analysis of Endosome Subpopulations of Rat Hepatocytes
3 E" t2 W; r5 @; } d14. Unit 3.12 Isolation of Synaptic Vesicles9 `/ ~6 t) R) C) ] x+ p6 }
15. Unit 3.13 Isolation of Clathrin-Coated Vesicles by Differential and Density Gradient Centrifugation
/ I, ^, k- j L) {/ z5 D16. Unit 3.14 Isolation of Melanosomes6 M* Q$ @2 g& J7 U: J6 H
17. Unit 3.15 Isolation of Lipid Droplets from Cells by Density Gradient Centrifugation
& w) ], j; Y+ w- p2 N18. Unit 3.16 Isolation of Mast Cell Granules
; ^9 Z: i2 c( C3 ]19. Unit 3.17 Immunoisolation of Centrosomes from Drosophila melanogaster4 J) l+ U' E" ]: S, P7 N. F
20. Unit 3.18 Isolation of Zymogen Granules from Rat Pancreas; {% u" m/ ~: i8 @1 e. n3 |. {* U
21. Unit 3.19 Isolation of Glyoxysomes from Pumpkin Cotyledons
) o% G: X' e9 \1 B; [22. Unit 3.20 Isolation of GLUT4 Storage Vesicles
7 {( D" N9 w/ `0 K, D2 M7 y: [23. Unit 3.21 Isolation of Intestinal Brush-Border Membranes) b: A2 s& h/ q& y& c
24. Unit 3.22 Isolation and Characterization of Exosomes from Cell Culture Supernatants and Biological
S7 ]3 l1 N, o' N3 cFluids* O! t; w: z+ v" o( _, w
25. Unit 3.23 Isolation of Intermediate Filaments
& u: Y$ K* ~, K- }. |+ i' S Y26. Unit 3.24 Isolation of T-Tubules from Skeletal Muscle
9 H# o! z, ~% C/ o27. Unit 3.25 Isolation of Myelin
* y* e& T# r: m, I. S4 O o28. Unit 3.26 Isolation of Renal Brush Borders! K& B! U$ s8 t- M# S
29. Unit 3.27 Isolation of Endoplasmic Reticulum, Mitochondria, and Mitochondria-Associated Membrane
8 e. M! o1 S& o, GFractions from Transfected Cells and from Human Cytomegalovirus-Infected Primary Fibroblasts
$ T$ u) d" F% M0 g9 X30. Unit 3.28 Isolation of Amyloplasts1 R) m5 F1 N& C# `
31. Unit 3.29 Isolation of Microtubules and Microtubule Proteins
! [) |( n9 r( R32. Unit 3.30 Purification of Intact Chloroplasts from Arabidopsis and Spinach Leaves by Isopycnic, h& F2 A9 Z4 n1 d1 d6 b
Centrifugation
& K' X" Z* T. Z. q5 [# } B33. Unit 3.31 Isolation of Neuromelanin Granules
Z. H e. a; L* `1 Q' B5 N9 R34. Unit 3.32 Isolation of Dense Core Secretory Vesicles from Pancreatic Endocrine Cells by Differential and
3 H4 [4 s0 j4 g4 t5 A4 BDensity Gradient Centrifugation
" S- j5 s: s5 {) i$ @35. Unit 3.33 Isolation and Biochemical Characterization of Amyloid Plaques and Paired Helical Filaments
# A5 A. s1 Z7 l5 z5 W36. Unit 3.34 Isolation of Legionella-Containing Vacuoles by Immuno-Magnetic Separation
4 ?3 E* q4 d, T) V. B, c" p37. Unit 3.35 Isolation of Platelet Granules
4 r6 c& u0 ^ v4 \5 w* u9 V7 z, {; w38. Unit 3.36 Isolation of Nucleoli t. @! H" |' [/ z7 \
39. Unit 3.37 Isolation of Cytotoxic T Cell and NK Granules and Purification of Their Effector Proteins8 H/ Y9 g8 O% y$ f; ]" U5 _
40. Unit 3.38 Isolation of Aggresomes and Other Large Aggregates
6 s5 t& w ~7 G0 Q41. Unit 3.39 Isolation of Chromaffin Granules) B9 W. }( o2 j$ t: l: L
42. Unit 3.40 Purification of Ribosomes from Human Cell Lines
# X/ k' ?* r. A: ]4 b1 r. E7 {6. Chapter 4 Microscopy
4 Y1 a' e4 U7 ?/ S1 D6 K1. Introduction/ E+ Y: T/ l3 `8 W% }
2. Unit 4.1 Proper Alignment and Adjustment of the Light Microscope
1 p- ^; P! `5 G$ F" w8 n3. Unit 4.2 Fluorescence Microscopy
7 A; @& ^( r: k/ o" p4. Unit 4.3 Immunofluorescence Staining* y' `3 \* m, M% f* f6 z. o
5. Unit 4.4 Fluorescent Staining of Subcellular Organelles: ER, Golgi Complex, and Mitochondria* [/ } r7 G0 P5 j3 r' ?# O K
6. Unit 4.5 Basic Confocal Microscopy
$ J$ M9 ]4 c" W+ J7. Unit 4.6 Immunoperoxidase Methods for Localization of Antigens in Cultured Cells and Tissues% m! T# l& _1 Z1 ]; Z
8. Unit 4.7 Cryo-Immunogold Electron Microscopy% K9 a7 l3 [" d& O- s+ x2 d3 o1 w- P
9. Unit 4.8 Correlative Video Light/Electron Microscopy
9 V6 Z( I% R) h' z. p- a10. Unit 4.9 Polarization Microscopy0 g5 k: N$ X2 y- j$ ] _. v0 B8 E R
11. Unit 4.10 Fluorescent Speckle Microscopy (FSM) of Microtubules and Actin in Living Cells
% @6 P% B ^% Z: b4 Z8 K12. Unit 4.11 Two-Photon Excitation Microscopy for the Study of Living Cells and Tissues
$ q& }; }! ?2 X j+ Q: b3 q13. Unit 4.12 Total Internal Reflection Fluorescence Microscopy for High-Resolution Imaging of Cell-Surface
8 G" S2 Q, p$ ^, t: KEvents
: T: |; L# F$ _; _14. Unit 4.13 Fluorescent Labeling of Yeast |! n: C5 U! Z) v+ z) M
15. Unit 4.14 Fluorescence Lifetime Imaging Microscopy
8 K- X# j; `! q. X3 Z16. Unit 4.15 Biological Second and Third Harmonic Generation Microscopy
+ `3 z) T. V( S8 X, L* E$ p17. Unit 4.16 Analyzing Real-Time Video Microscopy: The Dynamics and Geometry of Vesicles and Tubules) }" M1 m* R/ s% E2 K7 t+ z8 t
in Endocytosis) B- U! p u8 b1 m7 G6 j# L" m
18. Unit 4.17 Scanning Electron Microscopy of Cell Surface Morphology
+ \# y2 \& ]2 `! ^; L' @19. Unit 4.18 Fluorescence Imaging Techniques for Studying Drosophila Embryo Development* q# Q: F0 g6 ^2 x1 F
20. Unit 4.19 Quantitative Colocalization Analysis of Confocal Fluorescence Microscopy Images" ~6 _# e. k4 ?! t) q
21. Unit 4.20 Visualizing Protease Activity in Living Cells: From Two Dimensions to Four Dimensions
, r$ T1 r) O5 k1 ?22. Unit 4.21 Photoactivated Localization Microscopy (PALM) of Adhesion Complexes
( t9 f# {* ^& {- H23. Unit 4.22 Culturing MDCK Cells in Three Dimensions for Analyzing Intracellular Dynamics
5 \$ n. t5 X% l. P2 M" p+ o24. Unit 4.23 Interference Reflection Microscopy; Z6 [# w3 V1 l2 [+ Q
25. Unit 4.24 Fluorescence Correlation Spectroscopy in Living Cells: A Practical Approach
% i8 n, [! G* I26. Unit 4.25 Analysis of Mitochondrial Dynamics and Functions Using Imaging Approaches
& G( W k x9 B, P9 u27. Unit 4A Organelle Atlas: Appendix to Chapter 4
. ~3 n3 x4 p4 b Z6 x) t/ x7. Chapter 5 Characterization of Cellular Proteins9 i$ M& \; Z+ C- w$ @! ]
1. Introduction
' n/ O7 r9 h) ^2 w, g, n2. Unit 5.1 Overview of the Physical State of Proteins Within Cells
2 m: A) I* d7 p) u3. Unit 5.2 Determining the Topology of an Integral Membrane Protein
4 u! M( [& E2 ~' A1 o" h4. Unit 5.3 Determination of Molecular Size by Zonal Sedimentation Analysis on Sucrose Density Gradients
: ^5 B4 I" \. K( k0 d- D5. Unit 5.4 Analysis of the Association of Proteins with Membranes$ N/ T- k# b: g
6. Unit 5.5 Determination of Molecular Size by Size-Exclusion Chromatography (Gel Filtration)- }" q6 L% o7 D+ q: D _
7. Unit 5.6 Identification of Proteins in Complex Mixtures Using Liquid Chromatography and Mass
5 E; ?5 j5 _. p6 Q- C) R# GSpectrometry* [6 N( t4 u" a x2 [6 Q: Z: h
8. Unit 5.7 Determining Membrane Protein Topologies in Single Cells and High-Throughput Screening k$ K) c6 z9 z" P1 w
Applications$ C4 ?3 O- J3 e4 y
8. Chapter 6 Electrophoresis and Immunoblotting/ l) g! u& m1 p: T) `; V H
1. Introduction
8 S0 L# W' M" m2 ], \2. Unit 6.1 One-Dimensional SDS Gel Electrophoresis of Proteins
y4 I) W* [- l" i' g3. Unit 6.2 Immunoblotting and Immunodetection) Z8 M* y' h1 |# q2 u4 y4 X: T
4. Unit 6.3 Detection and Quantitation of Radiolabeled Proteins in Gels and Blots- R6 F) n2 P$ u4 m" E P( U
5. Unit 6.4 Two-Dimensional Gel Electrophoresis
% Q) }* j# U' i6. Unit 6.5 One-Dimensional Electrophoresis Using Nondenaturing Conditions# q R% R, Q( \7 ?; j
7. Unit 6.6 Staining Proteins in Gels
* `. `) t" D |# v: U8. Unit 6.7 Agarose Gel Electrophoresis of Proteins
9 {/ @4 ]8 Y. \- |" Z3 f4 V9. Unit 6.8 Fluorescence Detection of Glycoproteins in Gels and on Electroblots
9 P b, }, ]8 e: F& G D5 M10. Unit 6.9 Digital Electrophoresis Analysis
8 B8 l, O. }, Z: y! Y* T6 L11. Unit 6.10 Two-Dimensional Blue Native Polyacrylamide Gel Electrophoresis/ F: Q1 x. b/ h- e4 k" m
12. Unit 6.11 Measurement of Oxidatively-Induced Clustered DNA Lesions Using a Novel Adaptation of6 W9 O9 U3 X+ t, Z% a
Single Cell Gel Electrophoresis (Comet Assay)! H3 @+ [! H1 a$ ^1 y, d' s+ V
9. Chapter 7 Protein Labeling and Immunoprecipitation0 B; M5 [9 A! k1 b
1. Introduction
. G# ~3 ~. J5 v" h7 ?1 V2. Unit 7.1 Metabolic Labeling with Amino Acids
3 c3 F# Q* V- X3. Unit 7.2 Immunoprecipitation
2 E1 Z9 s a; _$ p' a3 I6 h% M1 W4. Unit 7.3 Metabolic Labeling with Sulfate8 _5 p9 \7 t, x
5. Unit 7.4 Metabolic Labeling with Fatty Acids) x3 u1 o. O5 t+ n2 r
6. Unit 7.5 Metabolic Labeling of Prenyl and Carboxyl-Methyl Groups' _ Q% ?. F. N# L
7. Unit 7.6 Metabolic Labeling and Immunoprecipitation of Yeast Proteins g; Q# u7 g2 h" `# J4 L
8. Unit 7.7 Metabolic Labeling and Immunoprecipitation of Drosophila Proteins
; d) ]1 T: h5 n3 L7 E9. Unit 7.8 Metabolic Labeling of Glycoproteins with Radioactive Sugars" z8 N4 ^- `0 A7 R
10. Unit 7.9 Analysis of Oxidative Modification of Proteins
8 z- ~1 J# J g4 L0 w6 Q11. Unit 7.10 Radioiodination of Cellular Proteins5 ~3 W8 g, k6 s; q% }. U) d
10. Chapter 8 Cell Cycle Analysis
) T2 ?, {+ h+ t5 F$ p1. Introduction9 A9 M% e3 d, \8 }
2. Unit 8.1 Overview of the Cell Cycle
1 q$ E3 B9 i& P$ `3. Unit 8.2 Assays for CDK Activity and DNA Replication in the Cell Cycle
; }( O& t' B1 E& J% m4. Unit 8.3 Methods for Synchronizing Cells at Specific Stages of the Cell Cycle
5 x- a" _+ n( R( T5. Unit 8.4 Determining Cell Cycle Stages by Flow Cytometry
% a0 y2 I, l, g3 O0 P+ a6. Unit 8.5 Centrifugal Elutriation to Obtain Synchronous Populations of Cells/ x" {( `: }' P2 c E( m
7. Unit 8.6 Dynamic Proliferation Assessment in Flow Cytometry
5 f/ [4 s+ \, L6 S j1 w) @11. Chapter 9 Cell Adhesion" k2 d: [; m, U# }
1. Introduction# B0 X. x; G, l2 t5 G9 N$ o
2. Unit 9.1 Cell-Substrate Adhesion Assays" a2 D3 w) ]% c, ^" ?- D2 i. Z
3. Unit 9.2 Quantitative Measurement of Cell Adhesion Using Centrifugal Force6 O; l/ e4 V# N, G K1 K
4. Unit 9.3 Cadherin-Dependent Cell-Cell Adhesion
# R8 x4 [, u9 n( D- [6 l5. Unit 9.4 Analyzing Integrin-Dependent Adhesion
; K& k* {" O( G/ [' B6. Unit 9.5 Analysis of Cell-Cell Contact Mediated by Ig Superfamily Cell Adhesion Molecules
9 }# C4 d* _6 i$ D7. Unit 9.6 Measurement of Adhesion Under Flow Conditions
/ U2 _3 W* D2 I F. e7 G12. Chapter 10 Extracellular Matrix3 r i; `" [$ p; O: A4 c
1. Introduction: a l" n* N6 [ K5 d7 Z8 V8 Q
2. Unit 10.1 Overview of Extracellular Matrix
2 i+ s t5 n. _/ ]# e3. Unit 10.2 Preparation of Basement Membrane Components from EHS Tumors) M) ^: Y( K ]) E- M0 D( Z7 T
4. Unit 10.3 Preparation of Gelled Substrates
" d! B& _6 P) e5. Unit 10.4 Preparation of Extracellular Matrices Produced by Cultured Corneal Endothelial and PF-HR97 A4 D5 f1 _" j* f
Endodermal Cells4 C& }* Z: h, i; W
6. Unit 10.5 Purification of Fibronectin
! q0 `7 A/ i( b# l2 l0 x7. Unit 10.6 Purification of Vitronectin
' e( J ?$ T% r& j8 T: _8. Unit 10.7 Proteoglycan Isolation and Analysis
/ M& q$ G+ f1 F" S9. Unit 10.8 Matrix Metalloproteinases7 \0 `+ b0 \2 F, H8 G n: b6 R
10. Unit 10.9 Preparation of Extracellular Matrices Produced by Cultured and Primary Fibroblasts9 R( C( y; M, [3 b/ {7 j/ {
11. Unit 10.10 Purification and Analysis of Thrombospondin-1' e0 l3 Y0 L) ]- D7 T( \
12. Unit 10.11 Purification of SPARC/Osteonectin( \2 E" T. K Z/ h3 h6 N# v5 m. F
13. Unit 10.12 Analysis of Fibronectin Matrix Assembly
' j0 Z6 _/ }. J$ Q8 [9 K& |% }14. Unit 10.13 Non-Radioactive Quantification of Fibronectin Matrix Assembly
9 Y9 ]+ W. U" N+ p& W9 }, n0 ?0 y0 |15. Unit 10.14 Use of Hyaluronan-Derived Hydrogels for Three-Dimensional Cell Culture and Tumor
5 Q4 u! p' o2 m. z$ `9 DXenografts
% W* E0 r5 m: A( h$ p! I; X+ l/ H16. Unit 10.15 Generation of Micropatterned Substrates Using Micro Photopatterning1 _9 y/ V$ B; {2 ^9 V
17. Unit 10.16 Preparation of Hydrogel Substrates with Tunable Mechanical Properties, j. m: ?1 W/ ]. g. Y7 w6 h
18. Unit 10.17 Engineering Three-Dimensional Collagen Matrices to Provide Contact Guidance during 3D5 Z) I( ?" ]: V: q4 T) \
Cell Migration
: E A* q5 M% q" E. d7 v7 r6 j19. Unit 10.18 Imaging Cells in Three-Dimensional Collagen Matrix
1 j, ~( a! f! Z13. Chapter 11 In Vitro Reconstitution
( @( r5 `- _: M( X$ u8 V* p1. Introduction4 T3 l3 j# }$ [* J9 b' G
2. Unit 11.1 Overview of Eukaryotic In Vitro Translation and Expression Systems; ~, n9 e+ A: E; o( s
3. Unit 11.2 In Vitro Translation
4 G- Y7 {5 s) Q4. Unit 11.3 In Vitro Analysis of Endoplasmic-Reticulum-to-Golgi Transport in Mammalian Cells9 a. ~% F9 ]6 p, |' i2 t" }
5. Unit 11.4 Cotranslational Translocation of Proteins into Canine Rough Microsomes+ R* E& l3 r8 u6 m, R
6. Unit 11.5 In Vitro Analysis of SV40 DNA Replication# |) g! I" ?2 P G! X P& |
7. Unit 11.6 In Vitro Transcription7 S) o: E1 z1 N" g: G3 r
8. Unit 11.7 Nuclear Import in Digitonin-Permeabilized Cells
5 H6 B3 y2 a g9. Unit 11.8 In Vitro Translation Using HeLa Extract2 Q* H% {6 C5 x8 e0 ]& x
10. Unit 11.9 Analysis of Eukaryotic Translation in Purified and Semipurified Systems* t2 u! e; c! n4 ~' U% h% w
11. Unit 11.10 Preparation and Use of Interphase Xenopus Egg Extracts- x) W% s+ F M1 C# ~1 F6 l: ?
12. Unit 11.11 Analysis of the Cell Cycle Using Xenopus Egg Extracts
0 g% h7 ?6 @. e: t0 J0 V2 R13. Unit 11.12 Analysis of Apoptosis Using Xenopus Egg Extracts
" w6 F% _1 ^: d( D+ T14. Unit 11.13 Mitotic Spindle Assembly In Vitro
7 {/ j! a2 ~5 Q/ u) L- F* p, u15. Unit 11.14 Analysis of RNA Export Using Xenopus Oocytes
& M3 L4 O/ ]: a+ Z, V* T% x16. Unit 11.15 In Vitro Analysis of Peroxisomal Protein Import) Y. `$ p5 h! h
17. Unit 11.16 In Vitro Analysis of Chloroplast Protein Import
! Z" W& U5 E/ p+ r" P18. Unit 11.17 In Vitro RNA Splicing in Mammalian Cell Extracts. g& G) Z8 Q& n/ X. h/ j! B+ a
19. Unit 11.18 Endocytosis Assays in Intact and Permeabilized Cells! K% B" C$ Z4 i4 U" Z/ I
20. Unit 11.19 In Vitro Analysis of Yeast Mitochondrial Protein Import5 G6 t9 @0 c b& I! Z
14. Chapter 12 Cell Motility
U- k; y4 ^0 }4 v. F( S1. Introduction
3 d/ s" F9 o. _& S2. Unit 12.1 Chemotaxis Assays for Eukaryotic Cells1 _% E1 x7 w1 \9 L
3. Unit 12.2 Invasion Assays
9 N$ `% M, y3 e3 O3 l5 D; U4. Unit 12.3 Cell Traction" C# o* U1 u' P8 u/ W+ G: e, {
5. Unit 12.4 Cell Wound Assays
9 I7 d: L7 X- Z" y6. Unit 12.5 Dictyostelium Cell Dynamics
, s, P7 c! _- B4 A d! b, H* k, Q7. Unit 12.6 Optical Microscopy.Based Migration Assay for Human Neutrophils4 L) P+ F# {% {4 z8 A
8. Unit 12.7 Actin-Based Motility Assay& r# ?7 W1 q# \" s
9. Unit 12.8 In Vivo Marking of Single Cells in Chick Embryos Using Photoactivation of GFP
2 s( j K8 o0 j- U7 Q, z. k9 o15. Chapter 13 Organelle Motility) S6 I) M6 m% \8 j
1. Introduction; L* ~! q! C; `7 @; B. J
2. Unit 13.1 Microtubule/Organelle Motility Assays
K; Y/ s' B1 C( S; j6 h/ O/ F3. Unit 13.2 In Vitro Motility Assay to Study Translocation of Actin by Myosin! K- ~! ? |; ^: d/ C/ x
4. Unit 13.3 Organelle Motility in Plant Cells: Imaging Golgi and ER Dynamics with GFP* q, c' {& X, T' C
5. Unit 13.4 Movement of Nuclei) e. B1 e* W& U+ ~: E) [7 a
6. Unit 13.5 Measuring Dynamics of Nuclear Proteins by Photobleaching7 Z0 v1 L |& g; O* q7 N
7. Unit 13.6 Functional Characterization of Proteins Regulating Actin Assembly
6 }, z7 u! m# i, ~7 Z' m Q! c16. Chapter 14 Signal Transduction: Protein Phosphorylation& z* S) j; ?. A- r9 Q4 M+ J
1. Introduction5 z+ r) L9 t) E! X, K, I8 J+ ]& h
2. Unit 14.1 Overview of Protein Phosphorylation8 v8 F* I% }( z" z* b) Q" m" l
3. Unit 14.2 Immunological Detection of Phosphorylation
3 h- S. b B. b# }' }1 p/ q3 [9 p4. Unit 14.3 The Detection of MAPK Signaling( h; x) ^0 }5 E
5. Unit 14.4 Labeling Cultured Cells with 32Pi and Preparing Cell Lysates for Immunoprecipitation
2 r* w8 |. z: ^5 ~3 m( p6. Unit 14.5 Phosphoamino Acid Analysis0 e0 P% e4 |7 H8 D7 y
7. Unit 14.6 Determination of Akt/PKB Signaling) T, {* l4 E( D8 t9 D
8. Unit 14.7 Analyzing FAK and Pyk2 in Early Integrin Signaling Events
* l2 U8 |: S8 N& ?6 X- z" n9. Unit 14.8 Rho GTPase Activation Assays
) A: a; c; q3 A1 A% t0 N) {: z( J10. Unit 14.9 In Vitro GEF and GAP Assays$ R$ S* F+ Z W. Y' U
11. Unit 14.10 In Vivo Imaging of Signal Transduction Cascades with Probes Based on Forster Resonance B3 d* m; |/ o4 K/ _
Energy Transfer (FRET)
* C0 J0 H2 O0 f# z' {12. Unit 14.11 Biosensors for Characterizing the Dynamics of Rho Family GTPases in Living Cells# k+ H! ?" @9 u) G1 I
13. Unit 14.12 Analysis of Arf GTP-Binding Protein Function in Cells2 J8 m& d6 x, C4 O1 {& Q% J
17. Chapter 15 Protein Trafficking
$ Q7 R$ z2 k8 f) q+ ^, R5 Q1. Introduction1 ~+ G$ K1 i1 D2 F3 M
2. Unit 15.1 Overview of Protein Trafficking in the Secretory and Endocytic Pathways
. l* R n" A$ v* x3 {8 W7 H8 D3. Unit 15.2 Use of Glycosidases to Study Protein Trafficking; d4 V6 C3 V% P% }' i& z, Y
4. Unit 15.3 Endocytosis: Biochemical Analyses; f; m8 g) R' M0 c# }" ?- O0 p' Z u
5. Unit 15.4 Determining Protein Transport to the Plasma Membrane' [# C! n z- ?% Q- Q- u& ?8 s+ |
6. Unit 15.5 Analysis of Membrane Traffic in Polarized Epithelial Cells
8 B' X d: X: y. @7. Unit 15.6 Analysis of Protein Folding and Oxidation in the Endoplasmic Reticulum
, O! S- L) s, i; f5 _* {4 P8. Unit 15.7 Measurements of Phagocytosis and Phagosomal Maturation
, B( D0 I$ D1 Z* R0 R) {% _, r. B9. Unit 15.8 Analysis of Protein Transport to Lysosomes1 K( u" j2 p1 ~ Q) S: W2 Z. `
10. Unit 15.9 Studies of the Ubiquitin Proteasome System
7 v- {9 B W6 e1 n5 z% L11. Unit 15.10 Measuring Retrograde Transport to the Trans-Golgi Network) k; }# q T! \! g4 }& x; C
12. Unit 15.11 Assays for Regulated Exocytosis of Mast Cell Granules
4 X6 l$ [' j4 W( X13. Unit 15.12 Analysis of Regulated Secretion Using PC12 Cells
9 t6 i0 g2 c) Y0 g14. Unit 15.13 Analysis of Endocytic Trafficking by Single-Cell Fluorescence Ratio Imaging- I* I; t8 ]9 {1 N: R" W' \
15. Unit 15.14 Quantitative Analysis of Endocytosis and Turnover of Epidermal Growth Factor (EGF) and& V1 T G, Z3 w$ v f& L
EGF Receptor
) P5 K/ p1 @5 @6 U" I3 K* |16. Unit 15.15 Documenting GLUT4 Exocytosis and Endocytosis in Muscle Cell Monolayers
, x% u/ s3 ?* v! o; @. ]18. Chapter 16 Antibodies as Cell Biological Tools: i6 N1 w' {* E9 x5 Y
1. Introduction
3 Z7 s! @, L$ d4 c% u; j8 R2. Unit 16.1 Production of Monoclonal Antibodies3 f3 M* t1 H$ Z; u* N3 S2 z8 d
3. Unit 16.2 Production of Polyclonal Antisera" H8 R: {; Q* Z
4. Unit 16.3 Purification of Immunoglobulin G
+ u' e/ }# i7 P* [ a5. Unit 16.4 Fragmentation of Immunoglobulin G( s+ `9 i: G/ x+ ?" {; i
6. Unit 16.5 Antibody Conjugates for Cell Biology6 R2 ?0 z& S$ I. c- M
7. Unit 16.6 Production of Antibodies That Recognize Specific Tyrosine-Phosphorylated Peptides
3 J1 ~1 ]5 C7 N: V19. Chapter 17 Macromolecular Interactions in Cells
5 c4 j( a7 j! [/ X( n8 G- S: M- |1. Introduction' Y9 L: ^2 q: |
2. Unit 17.1 Imaging Protein-Protein Interactions by Fluorescence Resonance Energy Transfer (FRET)2 w( g3 G/ g8 [7 V
Microscopy
/ s7 B6 J* y+ e3 Q, ?3. Unit 17.2 Identification of Protein Interactions by Far Western Analysis
+ G. z! O6 V/ _8 r7 B4. Unit 17.3 Interaction Trap/Two-Hybrid System to Identify Interacting Proteins7 i. _. ?0 a& d6 h) f
5. Unit 17.4 Mapping Protein-Protein Interactions with Phage-Displayed Combinatorial Peptide Libraries
) v# ~' [% t- j9 j9 O" U6. Unit 17.5 Protein-Protein Interactions Identified by Pull-Down Experiments and Mass Spectrometry
- w2 M9 _# h4 B. T7. Unit 17.6 Measuring Protein Interactions by Optical Biosensors
4 d/ v0 f6 a/ j7 |. C8. Unit 17.7 Chromatin Immunoprecipitation for Determining the Association of Proteins with Specific j1 z$ p1 E, m+ q) `' b
Genomic Sequences In Vivo
8 X0 p D6 p* k- j$ D; `9. Unit 17.8 Isothermal Titration Calorimetry! A- W# x+ ^! k- z
10. Unit 17.9 Rational Design and Evaluation of FRET Experiments to Measure Protein Proximities in Cells
) A" x# C" w6 e11. Unit 17.10 Identification and Analysis of Multiprotein Complexes Through Chemical Crosslinking
+ n1 S0 ?; F0 F12. Unit 17.11 Visualization of RNA Using Fluorescence Complementation Triggered by Aptamer-Protein
) i: z$ Z' e, W0 S4 qInteractions (RFAP) in Live Bacterial Cells$ N' U1 }; n) |. g: [
20. Chapter 18 Cellular Aging and Death2 E$ W2 @- D4 J1 y4 W/ ~3 U
1. Introduction3 \3 C* A$ R' {+ T4 E# Q5 W
2. Unit 18.1 Current Concepts in Cell Death7 v& ?, d8 R6 C* b
3. Unit 18.2 Analysis of Caspase Activation During Apoptosis4 g( x0 A1 M% ?' S5 T2 I
4. Unit 18.3 Assessment of Apoptosis and Necrosis by DNA Fragmentation and Morphological Criteria( I% \2 k- S: t; P* r
5. Unit 18.4 Quantitative Fluorescence In Situ Hybridization (Q-FISH)
! c& w$ f' Z$ w) y1 n9 t; g6. Unit 18.5 Analysis of Mitochondrial Dysfunction During Cell Death; z0 }5 o( X' b0 \5 o; N# k/ r+ t- G
7. Unit 18.6 Analysis of Telomeres and Telomerase
- n- m/ T) j ~, X8. Unit 18.7 Nonisotopic Methods for Determination of Poly(ADP-Ribose) Levels and Detection of. H$ D* f: v5 K2 @; ~
Poly(ADP-Ribose) Polymerase# J$ Y$ |. y& I, m4 S$ P9 W. m
9. Unit 18.8 Flow Cytometry of Apoptosis
8 Z t! ?' P0 s8 ]6 q* K10. Unit 18.9 Analysis of Cellular Senescence in Culture In Vivo: The Senescence-Associated -Galactosidase
9 l% T8 c$ C" D" h8 U z# dAssay
* E( j0 B/ l% K3 q11. Unit 18.10 High-Throughput Live Cell Imaging of Apoptosis
8 V# V; y+ p0 {- X- F. ^3 z8 {2 P21. Chapter 19 Whole Organism and Tissue Analysis
" c# _- l8 c. @2 \: P! Y( A/ x- v1. Introduction: B9 t }3 W/ |' y
2. Unit 19.1 Overview of Metastasis Assays u. J/ w& c, b0 Q4 X, N
3. Unit 19.2 Tail Vein Assay of Cancer Metastasis2 q) K: {: D2 w4 E0 |
4. Unit 19.3 Microanalysis of Gene Expression in Tissues Using T7-SAGE: Serial Analysis of Gene
6 k* n' U) C% u: s: o7 u" I9 CExpression After High-Fidelity T7-Based RNA Amplification) `' {* j: o( F/ o" B
5. Unit 19.4 SAGE Analysis from 1 兪g of Total RNA
O2 T/ r8 f. i( z) u- \) h( F; u6. Unit 19.5 The Chick Chorioallantoic Membrane as an In Vivo Angiogenesis Model4 f# S% r, w3 N
7. Unit 19.6 Experimental Metastasis Assays in the Chick Embryo
) e3 ?0 j8 q" c3 ~$ H8. Unit 19.7 Imaging Tumor Cell Movement In Vivo
! J0 }& E) U+ N1 d: {9 U9. Unit 19.8 Embryonic Organ Culture. Y0 n2 S J5 k, C- C6 J' K% m; O
10. Unit 19.9 Three-Dimensional Tissue Models of Normal and Diseased Skin
( ?- ?( a" E- F- ~ w+ [11. Unit 19.10 Overview: Engineering Transgenic Constructs and Mice
$ g2 \% e% M) y4 e1 l12. Unit 19.11 Generation of Transgenic Mice$ d$ R. j( D: x: f- M0 Z$ N- L
13. Unit 19.12 Overview: Generation of Gene Knockout Mice7 ?/ S/ o. j2 o3 g' e, Y/ p
14. Unit 19.13 Manipulation of Mouse Embryonic Stem Cells for Knockout Mouse Production4 X! {$ g5 j& U5 c+ z5 S
15. Unit 19.14 Generation of Gene Knockout Mice by ES Cell Microinjection
3 a' N Z: j8 Q22. Chapter 20 Expression and Introduction of Macromolecules into Cells+ e5 g: B& w% `% U5 f8 r& d8 @
1. Introduction& b6 k3 x4 c; R _
2. Unit 20.1 Direct Introduction of Molecules into Cells/ d( O9 o& `: Q4 T! A4 V
3. Unit 20.2 Protein Transduction: Generation of Full-Length Transducible Proteins Using the TAT System, G6 S( P% n _3 v" u
4. Unit 20.3 Calcium Phosphate Transfection
+ F' [8 ?8 ?# c0 M% E8 ~& F" i7 B5. Unit 20.4 Transfection Using DEAE-Dextran
) v0 o9 R( n+ Y! p0 G, _6. Unit 20.5 Transfection by Electroporation, w% B; s9 t E: O
7. Unit 20.6 Transfection of Cultured Eukaryotic Cells Using Cationic Lipid Reagents" g0 N) [# s) _% v0 ?, W# @
8. Unit 20.7 Optimization of Transfection
! v" Y( r4 u/ r6 X/ [( d8 P; \/ X3 G9. Unit 20.8 Inducible Gene Expression Using an Autoregulatory, Tetracycline-Controlled System
7 `5 S7 q" z' a: ?4 X4 z23. Chapter 21 Fluorescent Protein Technology
8 a* a* L& o q# R1. Introduction
* k# R n; K6 _6 u( z2. Unit 21.1 Measuring Protein Mobility by Photobleaching GFP Chimeras in Living Cells' E, r" h* O% P2 n1 _) _% ` @
3. Unit 21.2 Fluorescence Localization After Photobleaching (FLAP)
3 d3 R5 ?" @. v* |, {) x& s4. Unit 21.3 Visualization of Protein Interactions in Living Cells Using Bimolecular Fluorescence, Y" }% u* T% g W: j6 U
Complementation (BiFC) Analysis
# w# }: W, e9 X, p: k' h3 G5. Unit 21.4 Design and Use of Fluorescent Fusion Proteins in Cell Biology
* Q! ?- a- c; {4 r, r' Y6. Unit 21.5 The Fluorescent Protein Color Palette
( @/ C) I M. P5 h9 h; ~) Q7. Unit 21.6 Photoactivation and Imaging of Photoactivatable Fluorescent Proteins
H( A' \8 o; Z' c" P1 R24. Chapter 22 Cell Biology of Chromosomes and Nuclei: F) q y0 ]9 H( w3 S- D/ k
1. Introduction
+ R1 p1 I' P2 H3 @9 Q4 @% J D# [2. Unit 22.1 Overview of Cytogenetic Chromosome Analysis2 J3 d3 E) i! q" ]" @, Z
3. Unit 22.2 Preparation of Cytogenetic Specimens from Tissue Samples
5 N- y! t* B. G4. Unit 22.3 Traditional Banding of Chromosomes for Cytogenetic Analysis, k! L3 B. h( i8 _8 W& E. O
5. Unit 22.4 Fluorescence In Situ Hybridization (FISH)8 w8 i+ D* z# S j* x1 i. V+ U
6. Unit 22.5 Multi-Color FISH Techniques
: J4 [, N" b( F9 s) A. J6 e7. Unit 22.6 Comparative Genomic Hybridization
+ B% G2 Z8 s+ |' X: O: g; l1 x1 b. [8. Unit 22.7 Sister Chromatid Exchange! E0 f' o6 D9 E7 n
9. Unit 22.8 Detection of Mitotic Figures and Components of the Mitotic Machinery
' O; R% J3 L: c; z10. Unit 22.9 Assembly and Micromanipulation of Xenopus In Vitro.Assembled Mitotic Chromosomes
4 Z: A& h n8 o T% ]11. Unit 22.10 Replication Labeling with Halogenated Thymidine Analogs
- R6 q3 c1 B! a5 ^12. Unit 22.11 Assays for Ribosomal RNA Processing and Ribosome Assembly2 m( u: W/ c! L8 U
13. Unit 22.12 Visualization and Measurement of DNA Methyltransferase Activity in Living Cells
" l2 q2 c3 @5 R' o9 O14. Unit 22.13 Monitoring mRNA Export
: ^" m8 J k9 \15. Unit 22.14 Analysis of DNA Replication in Saccharomyces cerevisiae by Two-Dimensional and Pulsed-: A" E0 R5 a5 y# Y7 c6 k
Field Gel Electrophoresis
& x w' `) K- n, E# E9 G& a25. Chapter 23 Stem Cells* B- D" I* a3 _( k2 p
1. Introduction
) _7 B+ E. i1 w1 p0 t* g% V2. Unit 23.1 Stem Cells: An Overview8 n- B8 w6 p J/ _( I" N
3. Unit 23.2 Mouse Embryonic Stem Cell Derivation, and Mouse and Human Embryonic Stem Cell Culture1 J& d7 G: b( v6 W8 E
and Differentiation as Embryoid Bodies
- ?9 `' c' [: t4. Unit 23.3 Maintenance and In Vitro Differentiation of Mouse Embryonic Stem Cells to Form Blood
- F+ _0 u8 c* l( C/ o) jVessels7 {% |+ J' P9 {1 D; Y1 B0 H( A# f
5. Unit 23.4 Differentiation of Mouse Embryonic Stem Cells and of Human Adult Stem Cells into
6 o x$ `- q& q: h6 \Adipocytes
6 N# x* N' H. ]# ^6 G$ x6. Unit 23.5 Induction of ES Cell.Derived Cartilage Formation7 |" ]2 H G3 W |1 R# J, ^$ Q
7. Unit 23.6 Hematoendothelial Differentiation of Human Embryonic Stem Cells# v! V& P! C* s7 N
8. Unit 23.7 Neural Differentiation of Human ES Cells; e/ N5 d( [: Z* N3 d& k* Z8 I. m
26. Chapter 24 Lipids# r2 ?0 i2 c3 U/ g9 t% y4 M/ l
1. Introduction
" _$ O5 i7 c# |/ i0 y2. Unit 24.1 Using Fluorescent Sphingolipid Analogs to Study Intracellular Lipid Trafficking- q$ \* e5 y, S' k, c
3. Unit 24.2 Fluorescent Detection of Lipid Droplets and Associated Proteins0 a' m$ Q1 C: u' o0 d
4. Unit 24.3 Making Giant Unilamellar Vesicles via Hydration of a Lipid Film
, E4 n7 @1 A$ ]3 R+ l$ b5. Unit 24.4 Visualization of Cellular Phosphoinositide Pools with GFP-Fused Protein-Domains
/ `2 T" v+ c8 u# H9 n6 X" A27. Chapter 25 Nanotechnology
. `* s b- Y L1. Introduction: @4 @- [( ]/ M& s3 b2 e8 V1 W! M
2. Unit 25.1 In Vivo Imaging Using Quantum Dot.Conjugated Probes6 A9 g* b. ]8 F# r& m, r! ~
3. Unit 25.2 Fabrication and Application of Nanofibrous Scaffolds in Tissue Engineering( V9 `1 A& S6 w( M, j! x
28. Chapter 26 Viruses) d: k# B# E6 X' l1 ]
1. Introduction
! O; M$ o* U e; W9 l# t2. Unit 26.1 Production of Papillomavirus-Based Gene Transfer Vectors
' [+ w8 E# c. L( Z$ @+ b3. Unit 26.2 BK Virus (BKV): Infection, Propagation, Quantitation, Purification, Labeling, and Analysis of
+ e7 u2 M. |* ]1 k2 `" [. xCell Entry2 q" |8 X% g5 K2 [
4. Unit 26.3 Methods Used to Study Respiratory Virus Infection
, J8 T" U/ n0 U5. Unit 26.4 Compartmented Neuron Cultures for Directional Infection by Alpha Herpesviruses7 j! l6 G# q, S6 m9 e* F6 O# x( R
6. Unit 26.5 HIV-1 Interactions with Cells: From Viral Binding to Cell-Cell Transmission
3 {- l) A; f$ K5 z" Z4 ^; ~29. Chapter 26 Lipids
$ z4 J- Z& m9 G1. Unit 26.6 Methods for Monitoring Dynamics of Pulmonary RSV Replication by Viral Culture and by: B3 @! h% s) u9 X* ~2 B
Real-Time Reverse Transcription.PCR In Vivo: Detection of Abortive Viral Replication( Y* W8 x/ q' V" c/ V+ H
30. Chapter 27 RNA-Based Methods in Cell Biology @- P7 z L$ H h3 `3 D
1. Introduction
' e! O. W) s( }4 S S% ?5 A* b) l2. Unit 27.1 Silencing of Gene Expression in Cultured Cells Using Small Interfering RNAs- ~8 g5 B4 X. H( @: y
3. Unit 27.2 Gene Down-Regulation with Short Hairpin RNAs and Validation of Specificity by Inducible
: r" H# ^& e( W+ G2 z! e: M. A5 WRescue in Mammalian Cells; N: l1 h2 n* s6 S8 W4 q1 \) M
31. Appendix 1 Useful Information and Data; o4 e4 v1 L" N7 |. c
1. 1A Useful Measurements and Data+ b# A8 t# C7 m9 J
2. 1B Compendium of Drugs Commonly Used in Cell Biology Research0 i# |8 f& B! ^9 d2 F4 F G! V, R
3. 1C Identification of Motifs in Protein Sequences. X B/ p* s/ F7 W9 `0 E5 _5 w
4. 1D Safe Use of Radioisotopes) \' V/ h8 R) K# d! d
5. 1E Absorption and Emission Maxima for Common Fluorophores3 }. B8 ] V) X" U) \' j( V8 o
6. 1F Importing Biological Materials% m2 ]- D9 B2 m* G% h5 n
7. 1G Centrifuges and Rotors
- }' G' m2 ^; E8 F$ ?8. 1H Internet Basics for Biologists1 X k: E4 {" G/ w3 u" B
32. Appendix 2 Laboratory Stock Solutions and Equipment
5 v% y D0 s$ A1 j1. 2A Common Stock Solutions, Buffers, and Media
4 K2 h/ `) K6 Y: B7 C2. 2B Medium Formulations' Q0 @7 w. n. }7 w3 N$ E
3. 2C Standard Laboratory Equipment
+ z5 {2 A. E4 z( d6 O" h2 l1 {33. Appendix 3 Commonly Used Techniques* s) f3 Y5 N+ }, t4 _8 m- E: V1 j6 ~
1. 3A Molecular Biology Techniques
( x3 j3 ?5 n( R V2. 3B Spectrophotometric Determination of Protein Concentration
: U ^' u; }: T4 m. j3. 3C Dialysis and Concentration of Protein Solutions1 w) F: H/ _6 Q2 q
4. 3D Quantification of DNA and RNA with Absorption and Fluorescence Spectroscopy3 h2 d$ q ?7 R6 J4 A7 F
5. 3E Silanizing Glassware
; D3 ?( [% Z, W6. 3F Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization
1 x! O X4 X4 h8 E/ {. P; V7. 3G Micro RT-PCR
* y: A& l$ j1 h! Y8. 3H The Colorimetric Detection and Quantitation of Total Protein! n& n. q8 H" D3 Z
34. Appendix Suppliers
( @ h7 A% h& N- w! a3 B) T( _1. Selected Suppliers of Reagents and Equipment
; r2 H1 }% t' Q( ?! V( `# m7 b! W0 p6 {' y* u8 L0 Q( G m3 j' C2 Q
|
附件: 你需要登录才可以下载或查看附件。没有帐号？注册
-
总评分: 威望 + 20
包包 + 30
查看全部评分
|
发表于 2011-3-8 15:53
