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[干细胞与细胞生物学类] PDF电子书:Current Protocols in Cell Biology 2010版     [复制链接]

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本帖最后由 细胞海洋 于 2013-1-24 14:01 编辑   q3 ^! l- ~$ T, X! b
$ X9 m3 F+ R6 Z2 p: a
Current Protocols in Cell Biology 2010年完整版 5483页5 y" E( s* m+ y% o/ @
3 Y" y5 S+ W: n& c5 e
Online ISBN: 9780471143031
$ ?+ g& O  W% V' p7 C$ P* |5 C- eDOI: 10.1002/0471143030  m% v$ @7 f5 R

6 V3 N7 m! l& Z) `Table of Contents2 u0 @9 Z( m+ R
1. Preface
0 q" O, K3 H" m$ }" N6 g6 O2. Foreword
. v. Q1 B' }9 _" r/ p: y- B3 r2 H7 a4 w3. Chapter 1 Cell Culture7 L( q9 [3 j% `. z
1. Introduction
2 d; d  b: ~# y2. Unit 1.1 Basic Techniques in Mammalian Cell Tissue Culture; h) }& p& D5 t3 t
3. Unit 1.2 Media for Culture of Mammalian Cells2 \( R9 x4 [* G  F" n$ k# {3 F6 r6 d) O
4. Unit 1.3 Aseptic Technique for Cell Culture1 M* s! {$ U( R; r9 L
5. Unit 1.4 Sterilization and Filtration& S0 l2 ?' D' v) U+ [
6. Unit 1.5 Assessing and Controlling Microbial Contamination in Cell Cultures
" [  H: E: ]4 C% k0 d7. Unit 1.6 Media and Culture of Yeast& [7 g3 g) z6 n/ }- x* G* |2 v
8. Unit 1.7 BY-2 Cells: Culture and Transformation for Live Cell Imaging
2 q2 W  M: m; y% p' o) t4. Chapter 2 Preparation and Isolation of Cells. w% T; m, w6 D* @7 }
1. Introduction
1 a) R" [/ F$ c( V3 {) T2. Unit 2.1 Establishment of Fibroblast Cultures1 |; m% M! G5 G; n
3. Unit 2.2 Preparation and Culture of Human Lymphocytes
  c& x( [4 V& k% q* W$ b4. Unit 2.3 Preparation of Endothelial Cells0 u" i4 n+ Y$ W: W
5. Unit 2.4 Generation of Continuously Growing B Cell Lines by Epstein-Barr Virus Transformation
5 d! s# a- u: ^* @# R/ N% u6 k6. Unit 2.5 Laser Capture Microdissection
$ F* G( u# s" ?" z7. Unit 2.6 Preparation of Human Epidermal Keratinocyte Cultures2 g. M* ?7 g! a" J
8. Unit 2.7 Preparation and Coculture of Neurons and Glial Cells
) X1 {% R+ x. F2 B! i' S/ N0 F5. Chapter 3 Subcellular Fractionation and Isolation of Organelles
7 I" W5 o! y% f% n7 Z1. Introduction0 q2 J2 y" `# p  Q
2. Introduction
' t3 v# p2 L# I' C3. Unit 3.1 Overview of Cell Fractionation4 {* X& r* P9 w- u3 P0 w
4. Unit 3.2 Isolation of Rat Hepatocyte Plasma Membrane Sheets and Plasma Membrane Domains
. \9 u/ ]. M' I: _2 L$ j5. Unit 3.3 Isolation of Mitochondria from Tissues and Cells by Differential Centrifugation
) x% _- f8 T8 P! W: L! [6. Unit 3.4 Purification of a Crude Mitochondrial Fraction by Density-Gradient Centrifugation
/ w8 Q* U3 J& E4 q( `/ V" ~- D7. Unit 3.5 Isolation of Peroxisomes from Tissues and Cells by Differential and Density Gradient
( {4 Q4 D/ z5 [Centrifugation; _( f. g/ ~5 Y9 `* L; ~
8. Unit 3.6 Isolation of Lysosomes from Tissues and Cells by Differential and Density Gradient& b) l1 N! v+ \" w
Centrifugation
- G& z% Z5 x5 a: t/ V# }& z$ o6 t1 l( ~9. Unit 3.7 Overview of Subcellular Fractionation Procedures for the Yeast Saccharomyces cerevisiae
/ W8 h8 u) F+ f" H3 C8 p. s7 ?10. Unit 3.8 Isolation of Subcellular Fractions from the Yeast Saccharomyces cerevisiae$ x& F0 s, j( P1 v# r) C
11. Unit 3.9 Isolation of Golgi Membranes from Tissues and Cells by Differential and Density Gradient
& f0 s* ?3 S' B5 i' C4 i0 CCentrifugation
! N+ u3 W* M1 e9 l( T8 x  K12. Unit 3.10 Isolation of Nuclei and Nuclear Membranes From Animal Tissues" i. j9 K  h! R) c
13. Unit 3.11 Free-Flow Electrophoretic Analysis of Endosome Subpopulations of Rat Hepatocytes
/ w2 [/ [& o* ]  ~8 I/ M, ^14. Unit 3.12 Isolation of Synaptic Vesicles: {7 `1 r- k# ~" h. R3 n. W
15. Unit 3.13 Isolation of Clathrin-Coated Vesicles by Differential and Density Gradient Centrifugation2 L( Y6 H4 ~, [( a( p9 h2 `5 g5 A$ Q
16. Unit 3.14 Isolation of Melanosomes$ K6 Z5 t: t* H. [% L2 n3 l, ?
17. Unit 3.15 Isolation of Lipid Droplets from Cells by Density Gradient Centrifugation
4 w5 h+ z0 S8 \+ _& m18. Unit 3.16 Isolation of Mast Cell Granules
3 e1 n# {( @/ m3 I; O" Y0 N0 }19. Unit 3.17 Immunoisolation of Centrosomes from Drosophila melanogaster# ^7 j- Q0 A. J, _
20. Unit 3.18 Isolation of Zymogen Granules from Rat Pancreas
3 G: ^2 p  S) F21. Unit 3.19 Isolation of Glyoxysomes from Pumpkin Cotyledons# V7 K. p. q. }2 F
22. Unit 3.20 Isolation of GLUT4 Storage Vesicles
5 v" E" W0 h  R9 N2 @' K! s23. Unit 3.21 Isolation of Intestinal Brush-Border Membranes: ?4 t: k/ j' w* d, K
24. Unit 3.22 Isolation and Characterization of Exosomes from Cell Culture Supernatants and Biological4 @9 C$ X/ g9 y" X
Fluids
. n+ u) Y' C0 k, \( O25. Unit 3.23 Isolation of Intermediate Filaments
- ]0 H2 m" O0 G  C/ K, l4 k  J26. Unit 3.24 Isolation of T-Tubules from Skeletal Muscle- J9 o7 `) f, X! F
27. Unit 3.25 Isolation of Myelin
/ c: X( d( F1 Q2 \28. Unit 3.26 Isolation of Renal Brush Borders- g$ s2 L& T0 |. `' Y
29. Unit 3.27 Isolation of Endoplasmic Reticulum, Mitochondria, and Mitochondria-Associated Membrane
9 E( s. O6 P% [% S5 uFractions from Transfected Cells and from Human Cytomegalovirus-Infected Primary Fibroblasts* F) [5 }& E1 {7 B( a- ^9 u
30. Unit 3.28 Isolation of Amyloplasts
% }1 H# K- ~) b4 c2 S31. Unit 3.29 Isolation of Microtubules and Microtubule Proteins# ^7 o% R+ B' X( M! Z) b8 s
32. Unit 3.30 Purification of Intact Chloroplasts from Arabidopsis and Spinach Leaves by Isopycnic
: m8 M  `4 k& W  q" ~Centrifugation
# R. l4 A8 C& E  D) M, b- G33. Unit 3.31 Isolation of Neuromelanin Granules
. ~5 H- j* S. e2 ~34. Unit 3.32 Isolation of Dense Core Secretory Vesicles from Pancreatic Endocrine Cells by Differential and
* P! R4 a' V2 s. [' ?2 K! dDensity Gradient Centrifugation" u, p% Q6 v. R' a! J
35. Unit 3.33 Isolation and Biochemical Characterization of Amyloid Plaques and Paired Helical Filaments
* ?4 G" t3 Z8 u4 x, v% ]36. Unit 3.34 Isolation of Legionella-Containing Vacuoles by Immuno-Magnetic Separation
" I' V/ ]8 u/ A37. Unit 3.35 Isolation of Platelet Granules+ e2 d5 V# ]; v, X
38. Unit 3.36 Isolation of Nucleoli
2 B4 A3 i! A& y5 s39. Unit 3.37 Isolation of Cytotoxic T Cell and NK Granules and Purification of Their Effector Proteins0 ?7 t, E* H3 W( D5 [1 O6 H! I0 W
40. Unit 3.38 Isolation of Aggresomes and Other Large Aggregates; \; c) m2 Q/ [; R' x* R
41. Unit 3.39 Isolation of Chromaffin Granules4 s4 o$ `* O" X
42. Unit 3.40 Purification of Ribosomes from Human Cell Lines
/ h! T/ J  `. `" O6. Chapter 4 Microscopy: u) e0 t9 ~- \* x1 o2 {( E
1. Introduction7 \/ |$ V! N' x( F) w- v* k
2. Unit 4.1 Proper Alignment and Adjustment of the Light Microscope5 W9 S" C( \+ R, p. q
3. Unit 4.2 Fluorescence Microscopy
" {7 h! z$ ~% F# ?4 D9 l0 v4. Unit 4.3 Immunofluorescence Staining
4 j' y9 f/ c/ Q6 Y1 C$ t5. Unit 4.4 Fluorescent Staining of Subcellular Organelles: ER, Golgi Complex, and Mitochondria
5 L+ g. i4 h; D/ t6 K5 x, w6. Unit 4.5 Basic Confocal Microscopy( D* Z0 }; H7 @- c4 H" l7 s
7. Unit 4.6 Immunoperoxidase Methods for Localization of Antigens in Cultured Cells and Tissues+ q! e9 k' q  i' ~3 |( e5 P
8. Unit 4.7 Cryo-Immunogold Electron Microscopy
& N: x9 E2 B6 r5 R9. Unit 4.8 Correlative Video Light/Electron Microscopy
$ h9 r: a' V1 Q2 l) d6 y, F10. Unit 4.9 Polarization Microscopy5 I7 K3 x/ O9 r' K: K7 O
11. Unit 4.10 Fluorescent Speckle Microscopy (FSM) of Microtubules and Actin in Living Cells( ^8 x: ~, i% \1 O, e- x% ?
12. Unit 4.11 Two-Photon Excitation Microscopy for the Study of Living Cells and Tissues
8 y) {- _9 }/ N3 d+ ]13. Unit 4.12 Total Internal Reflection Fluorescence Microscopy for High-Resolution Imaging of Cell-Surface
# u+ a# R6 M0 n3 t! @1 UEvents
( w& u  `% g" K% s6 q& p14. Unit 4.13 Fluorescent Labeling of Yeast, H. l/ J$ I+ m6 h: y* N! r
15. Unit 4.14 Fluorescence Lifetime Imaging Microscopy
& F. W9 f% d  J2 F, o9 _16. Unit 4.15 Biological Second and Third Harmonic Generation Microscopy
6 K2 ~. Z2 T6 E( _5 o1 z17. Unit 4.16 Analyzing Real-Time Video Microscopy: The Dynamics and Geometry of Vesicles and Tubules: n/ `& ~/ B5 s/ e/ T
in Endocytosis
. R% O& ?0 s% s! s) ~18. Unit 4.17 Scanning Electron Microscopy of Cell Surface Morphology, S: t$ A% e# t* L
19. Unit 4.18 Fluorescence Imaging Techniques for Studying Drosophila Embryo Development
  j6 Z: }  ~" L7 T' [20. Unit 4.19 Quantitative Colocalization Analysis of Confocal Fluorescence Microscopy Images$ k4 f' S8 d6 }6 H5 x# z
21. Unit 4.20 Visualizing Protease Activity in Living Cells: From Two Dimensions to Four Dimensions5 h( S  u1 l. T
22. Unit 4.21 Photoactivated Localization Microscopy (PALM) of Adhesion Complexes
" ]$ D3 k+ ?* C+ P" s* d* [/ Q- |23. Unit 4.22 Culturing MDCK Cells in Three Dimensions for Analyzing Intracellular Dynamics
# o/ g6 N% }! s$ b, E- g# ]! X24. Unit 4.23 Interference Reflection Microscopy9 P( U% I4 F, o% p/ _
25. Unit 4.24 Fluorescence Correlation Spectroscopy in Living Cells: A Practical Approach3 ?. [; L1 K( |% I
26. Unit 4.25 Analysis of Mitochondrial Dynamics and Functions Using Imaging Approaches& a3 _7 f/ [; Y8 m% N
27. Unit 4A Organelle Atlas: Appendix to Chapter 4
3 G4 Y: _# b) R7. Chapter 5 Characterization of Cellular Proteins* {- {0 H  }. ?, w
1. Introduction
. T4 j$ r6 R; O$ L$ s) _2. Unit 5.1 Overview of the Physical State of Proteins Within Cells
5 J0 K2 e# C( r9 L5 ~$ ?3. Unit 5.2 Determining the Topology of an Integral Membrane Protein
, x( N4 u8 ]5 q' ?; q# o/ Y4. Unit 5.3 Determination of Molecular Size by Zonal Sedimentation Analysis on Sucrose Density Gradients
% K. _2 B8 H5 \5 a$ B/ k0 ^; m. M5. Unit 5.4 Analysis of the Association of Proteins with Membranes! e: L5 r9 q" p0 l, a
6. Unit 5.5 Determination of Molecular Size by Size-Exclusion Chromatography (Gel Filtration)
3 V0 t. j  f+ b/ |1 I9 b7. Unit 5.6 Identification of Proteins in Complex Mixtures Using Liquid Chromatography and Mass2 E/ R! Q; [$ i/ o" K( B
Spectrometry
; F: ]8 P" y7 h. t4 n9 U8. Unit 5.7 Determining Membrane Protein Topologies in Single Cells and High-Throughput Screening# \" G4 ^0 c4 Y& |
Applications
& a; M  \3 U- h8. Chapter 6 Electrophoresis and Immunoblotting
! M& I0 G& C  d  Z- d, {( }/ X1. Introduction
! ?6 [7 S4 ~' M2. Unit 6.1 One-Dimensional SDS Gel Electrophoresis of Proteins
1 g0 c. s. Q- @! O: U( o3. Unit 6.2 Immunoblotting and Immunodetection& y+ F1 L6 {' l
4. Unit 6.3 Detection and Quantitation of Radiolabeled Proteins in Gels and Blots% h' ]+ G/ O3 I, e1 J3 w
5. Unit 6.4 Two-Dimensional Gel Electrophoresis" f8 r' t! i- A4 y8 R% |6 D
6. Unit 6.5 One-Dimensional Electrophoresis Using Nondenaturing Conditions; D* X4 S  n& {- Q8 j% f
7. Unit 6.6 Staining Proteins in Gels( F. x- o$ {/ j2 a! [2 ^$ T
8. Unit 6.7 Agarose Gel Electrophoresis of Proteins* ]  {4 q9 T, r4 B6 ~$ K0 z
9. Unit 6.8 Fluorescence Detection of Glycoproteins in Gels and on Electroblots
1 ~& r7 t! N3 V: g3 W* e, z  I10. Unit 6.9 Digital Electrophoresis Analysis0 z  b6 I1 R% R/ C& _9 W
11. Unit 6.10 Two-Dimensional Blue Native Polyacrylamide Gel Electrophoresis
; ^8 E! w/ E9 J1 ~8 `# C12. Unit 6.11 Measurement of Oxidatively-Induced Clustered DNA Lesions Using a Novel Adaptation of; u% Z- U1 y6 V7 s" }. n4 ?
Single Cell Gel Electrophoresis (Comet Assay)! \0 d: H1 w. j: S7 d
9. Chapter 7 Protein Labeling and Immunoprecipitation' E: C1 s' [6 j5 O
1. Introduction8 F% u, j8 v0 T* h& ]
2. Unit 7.1 Metabolic Labeling with Amino Acids& c7 D( w8 l3 v% G# G
3. Unit 7.2 Immunoprecipitation
# ^; s7 o4 A1 l3 O4. Unit 7.3 Metabolic Labeling with Sulfate  n) U+ L& a: B" U# U; T
5. Unit 7.4 Metabolic Labeling with Fatty Acids& z6 ~0 f( J7 t* H3 \& N
6. Unit 7.5 Metabolic Labeling of Prenyl and Carboxyl-Methyl Groups
1 _: q$ w/ O+ ?8 `  M9 U7 X& V; H7. Unit 7.6 Metabolic Labeling and Immunoprecipitation of Yeast Proteins, T3 v$ R, p& b% S- Z( C- P( S! p
8. Unit 7.7 Metabolic Labeling and Immunoprecipitation of Drosophila Proteins
; ], k2 |6 ?6 M4 t9. Unit 7.8 Metabolic Labeling of Glycoproteins with Radioactive Sugars; I7 k# \' t8 I! g3 |% E
10. Unit 7.9 Analysis of Oxidative Modification of Proteins
! R: X6 M. {$ v% v5 Q9 G, a11. Unit 7.10 Radioiodination of Cellular Proteins- y, E8 a3 G, C. V0 ^
10. Chapter 8 Cell Cycle Analysis! `6 j4 s9 N3 x  m- p, b* [
1. Introduction* b9 D1 e) D6 t0 `
2. Unit 8.1 Overview of the Cell Cycle+ }3 X0 q) x. J$ Q7 i, z
3. Unit 8.2 Assays for CDK Activity and DNA Replication in the Cell Cycle5 Z  {$ c: b  t& m# f" S- o8 n9 p
4. Unit 8.3 Methods for Synchronizing Cells at Specific Stages of the Cell Cycle' \$ O3 \2 q: @4 K9 W# E+ V
5. Unit 8.4 Determining Cell Cycle Stages by Flow Cytometry" [4 [( y; F. y" }
6. Unit 8.5 Centrifugal Elutriation to Obtain Synchronous Populations of Cells
: ]9 |  \' F7 j' ?2 x+ D4 {7. Unit 8.6 Dynamic Proliferation Assessment in Flow Cytometry' x8 ]  M! ~2 ]+ {
11. Chapter 9 Cell Adhesion& H% L6 P: s' w3 Q' c
1. Introduction
1 u; v' X. M" F0 v! w$ U2. Unit 9.1 Cell-Substrate Adhesion Assays
+ I' D3 M! @8 C" L- K; V3. Unit 9.2 Quantitative Measurement of Cell Adhesion Using Centrifugal Force. R& x- U; X- s6 B, U+ B
4. Unit 9.3 Cadherin-Dependent Cell-Cell Adhesion
7 y8 F2 K4 `7 D" m9 D5. Unit 9.4 Analyzing Integrin-Dependent Adhesion. [  A/ x/ y( w* y4 ?1 v9 j8 {
6. Unit 9.5 Analysis of Cell-Cell Contact Mediated by Ig Superfamily Cell Adhesion Molecules
" O! d4 R( j1 {. _- A$ a* e7. Unit 9.6 Measurement of Adhesion Under Flow Conditions
2 }$ H2 J, \  x& |" j$ g12. Chapter 10 Extracellular Matrix
8 Q  f; H0 m- l* J1. Introduction
! y% Y: K3 h$ d% i) t  ~9 _# m8 R2 J2. Unit 10.1 Overview of Extracellular Matrix
" U7 U! G1 a# p3. Unit 10.2 Preparation of Basement Membrane Components from EHS Tumors+ w; t5 J6 l5 M/ z* B) G
4. Unit 10.3 Preparation of Gelled Substrates, c, ^- _5 y9 V  B- k
5. Unit 10.4 Preparation of Extracellular Matrices Produced by Cultured Corneal Endothelial and PF-HR96 o. n4 o- x  A1 r' u' k0 l. P
Endodermal Cells
, @" c' m# e$ i6. Unit 10.5 Purification of Fibronectin4 ~5 Z* R5 r9 Y) ^& b2 y
7. Unit 10.6 Purification of Vitronectin/ l, K) l- ?- e! P
8. Unit 10.7 Proteoglycan Isolation and Analysis
6 k$ m2 H! E+ ]* o( F$ J9. Unit 10.8 Matrix Metalloproteinases
, N) e  b$ A# Y/ a5 }0 f10. Unit 10.9 Preparation of Extracellular Matrices Produced by Cultured and Primary Fibroblasts( ^. z0 D& V$ u
11. Unit 10.10 Purification and Analysis of Thrombospondin-1- X6 U4 Z& j, P; P$ A
12. Unit 10.11 Purification of SPARC/Osteonectin
# h. Z) O9 y/ y* a! r  N13. Unit 10.12 Analysis of Fibronectin Matrix Assembly0 v$ H$ A+ e3 n7 {/ }. i
14. Unit 10.13 Non-Radioactive Quantification of Fibronectin Matrix Assembly8 r5 F% j# M' W
15. Unit 10.14 Use of Hyaluronan-Derived Hydrogels for Three-Dimensional Cell Culture and Tumor7 `7 a6 v3 U- j. K9 r& k
Xenografts
% R) _7 l' A5 m) ~! l# Z2 \" ^16. Unit 10.15 Generation of Micropatterned Substrates Using Micro Photopatterning
% W- S1 C* A1 F( P+ \9 |17. Unit 10.16 Preparation of Hydrogel Substrates with Tunable Mechanical Properties
, c( s4 l9 a# R18. Unit 10.17 Engineering Three-Dimensional Collagen Matrices to Provide Contact Guidance during 3D& r0 H# X+ T% i, p, V7 u5 v
Cell Migration, O' D. G! V9 I/ P- D% W
19. Unit 10.18 Imaging Cells in Three-Dimensional Collagen Matrix
. n# ]) M& G+ b3 I; M# K3 Q* i6 m( e13. Chapter 11 In Vitro Reconstitution
1 E; w9 q: j6 f' H1. Introduction9 C- J4 l0 a# T# C6 _( C' r+ i
2. Unit 11.1 Overview of Eukaryotic In Vitro Translation and Expression Systems1 J4 i2 Y. a; a, j  n4 I/ T
3. Unit 11.2 In Vitro Translation
% v0 H5 U- `0 |' f6 a4. Unit 11.3 In Vitro Analysis of Endoplasmic-Reticulum-to-Golgi Transport in Mammalian Cells; S( O9 g: b( _0 K
5. Unit 11.4 Cotranslational Translocation of Proteins into Canine Rough Microsomes
6 R1 [! [, y" _, A+ O4 |9 T) x7 K6. Unit 11.5 In Vitro Analysis of SV40 DNA Replication
9 x& o! u2 W5 Z% C5 k' z8 F7. Unit 11.6 In Vitro Transcription
( B7 {7 Z% D4 U/ i8. Unit 11.7 Nuclear Import in Digitonin-Permeabilized Cells8 D( u2 q! `4 A# g3 J
9. Unit 11.8 In Vitro Translation Using HeLa Extract' u2 }1 a; b* b$ G2 p& o
10. Unit 11.9 Analysis of Eukaryotic Translation in Purified and Semipurified Systems5 J( y1 f% Y  ~( e  A
11. Unit 11.10 Preparation and Use of Interphase Xenopus Egg Extracts4 z. E6 p7 Y9 q, W# y( I( Q0 I; s
12. Unit 11.11 Analysis of the Cell Cycle Using Xenopus Egg Extracts
2 r% y) y7 O6 e& k' o& Q13. Unit 11.12 Analysis of Apoptosis Using Xenopus Egg Extracts
' c2 {5 V9 Q5 t2 q14. Unit 11.13 Mitotic Spindle Assembly In Vitro
0 {  v4 z7 M- \1 L* |15. Unit 11.14 Analysis of RNA Export Using Xenopus Oocytes
! n0 Y5 o0 r- E# G; m$ ?3 S  G16. Unit 11.15 In Vitro Analysis of Peroxisomal Protein Import2 X8 B( w; V. L$ e! q9 `' r$ L
17. Unit 11.16 In Vitro Analysis of Chloroplast Protein Import
2 g4 E% F* X7 z8 O. w18. Unit 11.17 In Vitro RNA Splicing in Mammalian Cell Extracts! L& E, S  a6 Z! k1 r) p
19. Unit 11.18 Endocytosis Assays in Intact and Permeabilized Cells0 S, d5 K- \$ {9 s! Z' r; ^
20. Unit 11.19 In Vitro Analysis of Yeast Mitochondrial Protein Import
. A2 `6 z6 e, x14. Chapter 12 Cell Motility- b' L3 c4 r3 r) c, w% H1 @
1. Introduction9 [# @9 b: ?. H1 t
2. Unit 12.1 Chemotaxis Assays for Eukaryotic Cells4 x1 Z- o, b4 m, s. Q
3. Unit 12.2 Invasion Assays; w' R5 _! G8 N- s9 b( f: F
4. Unit 12.3 Cell Traction/ l* T7 P. m1 w' A3 ^! z) M0 C& q
5. Unit 12.4 Cell Wound Assays8 I* R6 u2 q4 R6 z6 [6 D
6. Unit 12.5 Dictyostelium Cell Dynamics' Y' K- K, k$ R  C7 s9 f& i% \
7. Unit 12.6 Optical Microscopy.Based Migration Assay for Human Neutrophils/ _3 t$ K0 O2 T: \
8. Unit 12.7 Actin-Based Motility Assay+ n( C) n- ~% Y1 R6 o
9. Unit 12.8 In Vivo Marking of Single Cells in Chick Embryos Using Photoactivation of GFP! Q! l% c% ]3 q/ b/ a  M8 k: O
15. Chapter 13 Organelle Motility
1 k6 n6 i) {3 }/ ]2 C! R0 r, i1. Introduction9 p- H1 N9 ?+ |& G. W5 X5 V+ v
2. Unit 13.1 Microtubule/Organelle Motility Assays' m2 Y3 z4 q' V6 m$ r) f
3. Unit 13.2 In Vitro Motility Assay to Study Translocation of Actin by Myosin( ?  F0 Z$ d/ P1 g, n" J" J
4. Unit 13.3 Organelle Motility in Plant Cells: Imaging Golgi and ER Dynamics with GFP
2 w3 |0 z5 ~0 v4 t  S/ U5. Unit 13.4 Movement of Nuclei% C; N2 |) ?5 A. `( h% C& n9 @
6. Unit 13.5 Measuring Dynamics of Nuclear Proteins by Photobleaching
. W5 f7 J  h0 _( i6 F: |7. Unit 13.6 Functional Characterization of Proteins Regulating Actin Assembly
$ H& M2 a- y( y0 K% ^4 b0 x* R16. Chapter 14 Signal Transduction: Protein Phosphorylation
9 f$ W- U( Z3 ?+ O% c  \4 @1. Introduction% i+ I, Z: `  \! P
2. Unit 14.1 Overview of Protein Phosphorylation
! z7 y2 R1 V$ z3 u% |. w4 \& h3. Unit 14.2 Immunological Detection of Phosphorylation
+ f1 \4 y6 D' W+ U4. Unit 14.3 The Detection of MAPK Signaling
% L  [0 U  |3 w0 \: ~' f! _6 K& G5. Unit 14.4 Labeling Cultured Cells with 32Pi and Preparing Cell Lysates for Immunoprecipitation
" x0 @+ D% o4 S/ L4 G3 L2 I6. Unit 14.5 Phosphoamino Acid Analysis
8 }* x5 }. A) k5 Y9 ]6 R5 `7. Unit 14.6 Determination of Akt/PKB Signaling- N) T3 o9 ^6 F9 L7 N- W! P# H
8. Unit 14.7 Analyzing FAK and Pyk2 in Early Integrin Signaling Events
5 _- ]" j' V  S/ b9. Unit 14.8 Rho GTPase Activation Assays
5 i6 S& T( k) \" @$ M; F10. Unit 14.9 In Vitro GEF and GAP Assays
) ~8 y& O; j7 c6 {' w8 T2 h11. Unit 14.10 In Vivo Imaging of Signal Transduction Cascades with Probes Based on Forster Resonance/ g5 R2 f( |" {, w2 U
Energy Transfer (FRET)/ O9 m9 t( Q  z: q6 @0 k
12. Unit 14.11 Biosensors for Characterizing the Dynamics of Rho Family GTPases in Living Cells5 [0 _* e8 z% X' {( P
13. Unit 14.12 Analysis of Arf GTP-Binding Protein Function in Cells, O: }8 n. p# B8 @7 L' l# _4 t$ k1 i$ t
17. Chapter 15 Protein Trafficking; J1 c  d% S& n- i  J
1. Introduction9 C: \- c" ~) x/ |3 R+ S" e
2. Unit 15.1 Overview of Protein Trafficking in the Secretory and Endocytic Pathways
: N, ?- a2 N4 s/ [3. Unit 15.2 Use of Glycosidases to Study Protein Trafficking
" f/ A) h( i; m( |4. Unit 15.3 Endocytosis: Biochemical Analyses7 c0 l) X$ l% }  e
5. Unit 15.4 Determining Protein Transport to the Plasma Membrane
' `# Z4 v4 a. G9 y9 R6. Unit 15.5 Analysis of Membrane Traffic in Polarized Epithelial Cells
9 E, Z! P! f2 u' K7. Unit 15.6 Analysis of Protein Folding and Oxidation in the Endoplasmic Reticulum
7 t$ @8 J, n) T/ C" E# v( s; q5 V8. Unit 15.7 Measurements of Phagocytosis and Phagosomal Maturation- D% A; z  g' X
9. Unit 15.8 Analysis of Protein Transport to Lysosomes# F, S* g+ l4 Z1 R
10. Unit 15.9 Studies of the Ubiquitin Proteasome System
  n7 V% X6 ?( u! f11. Unit 15.10 Measuring Retrograde Transport to the Trans-Golgi Network
/ Q( `: W. s# s2 t  e  H12. Unit 15.11 Assays for Regulated Exocytosis of Mast Cell Granules
  m0 V0 O& n: A13. Unit 15.12 Analysis of Regulated Secretion Using PC12 Cells; ?& w: s: V3 {* I) ]+ G( G
14. Unit 15.13 Analysis of Endocytic Trafficking by Single-Cell Fluorescence Ratio Imaging6 P+ O% T+ v: A6 {+ o
15. Unit 15.14 Quantitative Analysis of Endocytosis and Turnover of Epidermal Growth Factor (EGF) and0 n* m* F+ ^. G; T" J1 C) j
EGF Receptor* U) f% l6 G( f+ Q
16. Unit 15.15 Documenting GLUT4 Exocytosis and Endocytosis in Muscle Cell Monolayers: ^3 A" y* ]2 M6 v2 r* |
18. Chapter 16 Antibodies as Cell Biological Tools0 \! W: u1 j" _1 `9 v
1. Introduction
$ H" `4 ~* [$ k+ N6 }8 t: Y7 k2. Unit 16.1 Production of Monoclonal Antibodies
) o4 ~+ w' [1 R6 V% d2 r3. Unit 16.2 Production of Polyclonal Antisera
, h/ f  y  w+ g6 r, M4. Unit 16.3 Purification of Immunoglobulin G! Z1 a+ Q- p: D+ k4 D! i2 p9 ~4 s
5. Unit 16.4 Fragmentation of Immunoglobulin G' P* v; i( i7 I0 O! a9 M" D' n3 h
6. Unit 16.5 Antibody Conjugates for Cell Biology( }$ `# Z2 E) _/ `
7. Unit 16.6 Production of Antibodies That Recognize Specific Tyrosine-Phosphorylated Peptides
' ?* X% a  t! q  A19. Chapter 17 Macromolecular Interactions in Cells
8 _+ Y/ f* Z; o& b: l4 V1. Introduction; G8 b) T, r* I/ s0 U: l
2. Unit 17.1 Imaging Protein-Protein Interactions by Fluorescence Resonance Energy Transfer (FRET)4 q) O* f- h% o0 J7 Y
Microscopy
* C' b6 Y4 _* S" K# u3. Unit 17.2 Identification of Protein Interactions by Far Western Analysis
* |8 Q9 |4 u! x7 j4. Unit 17.3 Interaction Trap/Two-Hybrid System to Identify Interacting Proteins- V! @4 |) J- [# o0 U2 K
5. Unit 17.4 Mapping Protein-Protein Interactions with Phage-Displayed Combinatorial Peptide Libraries
& W! W+ K  D1 ^. y+ z8 s! e6. Unit 17.5 Protein-Protein Interactions Identified by Pull-Down Experiments and Mass Spectrometry8 J5 W0 \1 O, r' D0 C; g
7. Unit 17.6 Measuring Protein Interactions by Optical Biosensors* F3 @/ t6 V  A7 d( {1 D7 G
8. Unit 17.7 Chromatin Immunoprecipitation for Determining the Association of Proteins with Specific6 E5 d& @. C( _8 a+ e
Genomic Sequences In Vivo/ t0 F& o" k; P( I4 x5 I
9. Unit 17.8 Isothermal Titration Calorimetry
9 L" V$ d# l6 ?: f+ k  Z% s10. Unit 17.9 Rational Design and Evaluation of FRET Experiments to Measure Protein Proximities in Cells
4 B4 }7 D8 N8 L11. Unit 17.10 Identification and Analysis of Multiprotein Complexes Through Chemical Crosslinking
& W4 [1 e+ {, B0 r! s" ?6 C$ k12. Unit 17.11 Visualization of RNA Using Fluorescence Complementation Triggered by Aptamer-Protein1 p& `8 I6 z7 t" z! ?
Interactions (RFAP) in Live Bacterial Cells
# F: f4 {6 v! P9 }+ M20. Chapter 18 Cellular Aging and Death' I. O8 C% s! {* ]/ Q
1. Introduction
: I# O3 a1 ]. y3 a4 M2. Unit 18.1 Current Concepts in Cell Death
$ g  [. V; j. z. H3. Unit 18.2 Analysis of Caspase Activation During Apoptosis
  B! t4 R" {  q( C0 g- r9 M4. Unit 18.3 Assessment of Apoptosis and Necrosis by DNA Fragmentation and Morphological Criteria
' R% f2 q& m+ F0 f! X. O5. Unit 18.4 Quantitative Fluorescence In Situ Hybridization (Q-FISH)! u2 s. z9 L6 N( u
6. Unit 18.5 Analysis of Mitochondrial Dysfunction During Cell Death3 @( r/ e) H5 d' @3 d, T( x
7. Unit 18.6 Analysis of Telomeres and Telomerase
( b% E% X! r0 ?5 Q7 R+ v( ]8. Unit 18.7 Nonisotopic Methods for Determination of Poly(ADP-Ribose) Levels and Detection of
9 D. y0 P+ R$ i% \Poly(ADP-Ribose) Polymerase4 n# n+ W% M1 h# c( \
9. Unit 18.8 Flow Cytometry of Apoptosis9 `8 ?& f* {& J
10. Unit 18.9 Analysis of Cellular Senescence in Culture In Vivo: The Senescence-Associated -Galactosidase
) w' o. f- K+ I# O& P# D3 i# {Assay% {- n: u- W) G  u( G9 j) t
11. Unit 18.10 High-Throughput Live Cell Imaging of Apoptosis
" e' s, F& `* s21. Chapter 19 Whole Organism and Tissue Analysis- e; @% Z3 D9 v( c5 Z; R' F- h
1. Introduction
8 b2 P$ ]4 [5 b2. Unit 19.1 Overview of Metastasis Assays. [$ l. l  E2 A0 b1 ]8 t
3. Unit 19.2 Tail Vein Assay of Cancer Metastasis1 ~# v5 }% F% S2 L7 p' z' G6 B
4. Unit 19.3 Microanalysis of Gene Expression in Tissues Using T7-SAGE: Serial Analysis of Gene
! y  Y9 b* l7 o8 H. s& U( ^/ xExpression After High-Fidelity T7-Based RNA Amplification
% x$ K- o( F- n- h5. Unit 19.4 SAGE Analysis from 1 兪g of Total RNA4 E3 F6 E+ H: }0 T( J( D8 N* c
6. Unit 19.5 The Chick Chorioallantoic Membrane as an In Vivo Angiogenesis Model
" C) e% t  e$ F% B: w( y! s; a7. Unit 19.6 Experimental Metastasis Assays in the Chick Embryo
. \1 f. h; P. i1 }3 r1 U0 @8. Unit 19.7 Imaging Tumor Cell Movement In Vivo, F4 H% P6 W3 j' @
9. Unit 19.8 Embryonic Organ Culture7 ?4 u* A7 B' W
10. Unit 19.9 Three-Dimensional Tissue Models of Normal and Diseased Skin& w5 b1 N  O2 x+ b% X, p* m6 r
11. Unit 19.10 Overview: Engineering Transgenic Constructs and Mice
6 u+ }0 p( P. {' ?! @$ B& P12. Unit 19.11 Generation of Transgenic Mice6 X' h. y% h" P! c% ?; ~" L# b
13. Unit 19.12 Overview: Generation of Gene Knockout Mice1 s" o3 O  X: B9 r
14. Unit 19.13 Manipulation of Mouse Embryonic Stem Cells for Knockout Mouse Production
6 p6 T# S8 _* K8 h0 O8 H& S' k* \15. Unit 19.14 Generation of Gene Knockout Mice by ES Cell Microinjection
. z# L# k0 \1 `+ C( J( Z2 [22. Chapter 20 Expression and Introduction of Macromolecules into Cells; Y( V5 A* s' V& l" ~7 Q" |( U
1. Introduction
" |0 c. g, x1 ]2. Unit 20.1 Direct Introduction of Molecules into Cells
& i4 f. \# r* U9 W3. Unit 20.2 Protein Transduction: Generation of Full-Length Transducible Proteins Using the TAT System
4 P1 {; ~( `8 x% ?) K4. Unit 20.3 Calcium Phosphate Transfection
8 D  I% S$ O$ h' `2 {, _5. Unit 20.4 Transfection Using DEAE-Dextran+ L! p- C6 K6 Q, z; T
6. Unit 20.5 Transfection by Electroporation
* Y2 N# a  ]8 U! Q. f! p. t9 V7. Unit 20.6 Transfection of Cultured Eukaryotic Cells Using Cationic Lipid Reagents, b2 o0 k, x$ h& \8 E& w
8. Unit 20.7 Optimization of Transfection& s0 c5 H' C) b) c' \0 S
9. Unit 20.8 Inducible Gene Expression Using an Autoregulatory, Tetracycline-Controlled System( e* B" ]+ U5 _; L5 E4 h
23. Chapter 21 Fluorescent Protein Technology& Z5 t4 L8 {* o
1. Introduction; W  d7 c9 ]8 C/ B; h/ Y) P. w
2. Unit 21.1 Measuring Protein Mobility by Photobleaching GFP Chimeras in Living Cells' o' E* i8 \* a; M, D' m7 m; @: d
3. Unit 21.2 Fluorescence Localization After Photobleaching (FLAP), F- T* [+ Y4 x4 j7 ?# k" ~% r, p4 R
4. Unit 21.3 Visualization of Protein Interactions in Living Cells Using Bimolecular Fluorescence' Z) h: u' ~9 ^4 n; P9 f
Complementation (BiFC) Analysis5 H1 c0 z0 h( d" ]+ E
5. Unit 21.4 Design and Use of Fluorescent Fusion Proteins in Cell Biology
% e* `6 x, N) `6. Unit 21.5 The Fluorescent Protein Color Palette" d/ Q, }6 D+ e' g
7. Unit 21.6 Photoactivation and Imaging of Photoactivatable Fluorescent Proteins$ |. X2 U* H- h% V: T4 H" A9 L
24. Chapter 22 Cell Biology of Chromosomes and Nuclei
9 `5 M. T- B7 k* p- r4 {# V: E1. Introduction6 w& o% S2 @; Q
2. Unit 22.1 Overview of Cytogenetic Chromosome Analysis9 e, X$ l; i' h$ _6 g6 f9 f
3. Unit 22.2 Preparation of Cytogenetic Specimens from Tissue Samples5 k! E! }8 ^- G& n( B) y
4. Unit 22.3 Traditional Banding of Chromosomes for Cytogenetic Analysis: D9 Q) p5 m+ q( f; y/ [
5. Unit 22.4 Fluorescence In Situ Hybridization (FISH)/ @* \/ t7 ?% d2 r
6. Unit 22.5 Multi-Color FISH Techniques
; p/ a. l: W1 F6 O7. Unit 22.6 Comparative Genomic Hybridization
" u# `. y+ m/ c7 U8. Unit 22.7 Sister Chromatid Exchange
0 @& M% @! s) K* @9. Unit 22.8 Detection of Mitotic Figures and Components of the Mitotic Machinery
; T  V9 q4 V" v8 R10. Unit 22.9 Assembly and Micromanipulation of Xenopus In Vitro.Assembled Mitotic Chromosomes
, {9 z0 i8 R, f5 k- C11. Unit 22.10 Replication Labeling with Halogenated Thymidine Analogs
7 z4 p; c7 F4 j7 A8 z: ]( N" Q12. Unit 22.11 Assays for Ribosomal RNA Processing and Ribosome Assembly
" T. W+ T" q4 R4 V( ?4 |13. Unit 22.12 Visualization and Measurement of DNA Methyltransferase Activity in Living Cells) m. ^- z) j7 N# [  u& o* a( U
14. Unit 22.13 Monitoring mRNA Export8 i- e4 |9 j3 w2 a$ \. Z
15. Unit 22.14 Analysis of DNA Replication in Saccharomyces cerevisiae by Two-Dimensional and Pulsed-3 S: P% U$ {. A/ n+ G; e2 a9 O$ q+ G
Field Gel Electrophoresis
/ I* `1 j$ M! `. ~5 p25. Chapter 23 Stem Cells
8 o) \* z$ ?  |! L! V. y; n1. Introduction4 h% {) a  e* r! q9 q2 b- S
2. Unit 23.1 Stem Cells: An Overview
2 r! ?  _' f  S' b0 G4 u. L3. Unit 23.2 Mouse Embryonic Stem Cell Derivation, and Mouse and Human Embryonic Stem Cell Culture
3 b' H. S& K1 u* Qand Differentiation as Embryoid Bodies5 [& P3 p. U" `
4. Unit 23.3 Maintenance and In Vitro Differentiation of Mouse Embryonic Stem Cells to Form Blood3 _- |8 Q) \: x; V. i# Q
Vessels
* r* d4 C- m. J% L! W9 _( M* L) m3 {6 |5. Unit 23.4 Differentiation of Mouse Embryonic Stem Cells and of Human Adult Stem Cells into
! r' [6 N. x& J7 p( V& D% H# @Adipocytes" T6 z! J8 ~4 D+ k% D. h. a3 k  Y
6. Unit 23.5 Induction of ES Cell.Derived Cartilage Formation4 T+ r8 k1 [% O: i
7. Unit 23.6 Hematoendothelial Differentiation of Human Embryonic Stem Cells& b! q2 q1 Z# ?5 z# J2 w
8. Unit 23.7 Neural Differentiation of Human ES Cells) f/ B/ H- m) {$ q
26. Chapter 24 Lipids2 q8 o! E1 N1 T- Y; n  C9 B
1. Introduction% r! \) r3 t+ i6 ?5 g6 [6 X* O, q- h2 v
2. Unit 24.1 Using Fluorescent Sphingolipid Analogs to Study Intracellular Lipid Trafficking/ r$ N  k+ Q# M5 y
3. Unit 24.2 Fluorescent Detection of Lipid Droplets and Associated Proteins0 K  s! G" P6 }- |3 s
4. Unit 24.3 Making Giant Unilamellar Vesicles via Hydration of a Lipid Film/ a. [8 ^$ t3 r1 b# E; T) O, p) {
5. Unit 24.4 Visualization of Cellular Phosphoinositide Pools with GFP-Fused Protein-Domains
9 p0 V5 k: O, _$ ?/ P7 V$ T27. Chapter 25 Nanotechnology6 I: [$ W3 e: n) r  V9 D  A
1. Introduction
* s; f% n* ]& A; d2. Unit 25.1 In Vivo Imaging Using Quantum Dot.Conjugated Probes
6 Y, }* K$ Z- V. o5 c- M, L% n9 N3. Unit 25.2 Fabrication and Application of Nanofibrous Scaffolds in Tissue Engineering# d. ?/ m5 a2 {
28. Chapter 26 Viruses' {) A) K" p7 n  Z: A; B" ~
1. Introduction* E1 P7 [$ S" L8 Z3 P$ M
2. Unit 26.1 Production of Papillomavirus-Based Gene Transfer Vectors; s! a7 B( x/ x
3. Unit 26.2 BK Virus (BKV): Infection, Propagation, Quantitation, Purification, Labeling, and Analysis of
: [8 P4 [7 `+ @2 BCell Entry; N  e5 l, [; u% o! s) P
4. Unit 26.3 Methods Used to Study Respiratory Virus Infection
; T) q; P1 ^: V8 f% X' y1 ~# T5. Unit 26.4 Compartmented Neuron Cultures for Directional Infection by Alpha Herpesviruses$ ]1 ]8 d7 i* u4 m+ p2 g; d& V
6. Unit 26.5 HIV-1 Interactions with Cells: From Viral Binding to Cell-Cell Transmission
7 y1 }$ e8 ^. `  M1 B( a9 O29. Chapter 26 Lipids
5 A3 v, m9 O8 |1 n( B1. Unit 26.6 Methods for Monitoring Dynamics of Pulmonary RSV Replication by Viral Culture and by$ O' s$ r! F, b
Real-Time Reverse Transcription.PCR In Vivo: Detection of Abortive Viral Replication
% @& e7 I+ R$ S+ Q* o8 g% e30. Chapter 27 RNA-Based Methods in Cell Biology
- a0 h- w: ]. w% i: B) R1 F1. Introduction3 {: D* ^3 d  h- y) \
2. Unit 27.1 Silencing of Gene Expression in Cultured Cells Using Small Interfering RNAs0 i1 b. ~" u# v( @4 f- O5 H/ }8 S
3. Unit 27.2 Gene Down-Regulation with Short Hairpin RNAs and Validation of Specificity by Inducible
: v/ a' f  N; D* X' [Rescue in Mammalian Cells! r  |8 C( @' P2 [# i- s
31. Appendix 1 Useful Information and Data
8 b( i+ s" }1 x$ O5 }- p1. 1A Useful Measurements and Data
" h0 _. L/ J; w2. 1B Compendium of Drugs Commonly Used in Cell Biology Research
1 @& B5 _' G0 x7 H+ Q3. 1C Identification of Motifs in Protein Sequences
2 ~: w) s1 [" f( W2 j9 B/ j6 l4. 1D Safe Use of Radioisotopes
; g+ P' h: u( q6 k5. 1E Absorption and Emission Maxima for Common Fluorophores
4 ^4 X, j$ G+ r0 G+ w6. 1F Importing Biological Materials  C/ R4 P) `5 l3 S" i
7. 1G Centrifuges and Rotors
" Y8 U; U- c" y& X5 c3 e8. 1H Internet Basics for Biologists
4 z" m% a7 ~* C32. Appendix 2 Laboratory Stock Solutions and Equipment' A+ ]. P& ?; K( q9 v4 l/ {' P
1. 2A Common Stock Solutions, Buffers, and Media: G/ \* H  U+ d$ @( m
2. 2B Medium Formulations
: ~9 N: ~% ?# P, p( I! y; ?3. 2C Standard Laboratory Equipment
" d% L7 j- x) n( m$ z33. Appendix 3 Commonly Used Techniques
* ]! w' A# e6 e1. 3A Molecular Biology Techniques
4 k4 H/ x- I( D  Z0 \& h: ^2. 3B Spectrophotometric Determination of Protein Concentration
: Q( o8 V( e: F- x) C9 W, J7 U3. 3C Dialysis and Concentration of Protein Solutions
8 J5 N) \, t! r1 V) F4. 3D Quantification of DNA and RNA with Absorption and Fluorescence Spectroscopy
2 t% Y) d& l6 c) C* [" q7 `5. 3E Silanizing Glassware% B9 w% I/ B7 p" F# `& z6 W
6. 3F Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization
& |# r0 H# N% t/ W7. 3G Micro RT-PCR, j4 g+ l  w0 s& }0 g4 N
8. 3H The Colorimetric Detection and Quantitation of Total Protein
, b* P. d# i/ h# G34. Appendix Suppliers* |# K" H) ~4 X) H; |; {) x/ d" G
1. Selected Suppliers of Reagents and Equipment: h$ D) x+ V7 A$ Z& c/ x

" _* Q$ J  o+ i5 u& l7 E: i
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沙发
发表于 2011-3-8 18:58 |只看该作者
taiguile

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好东西,谢谢+ h+ Z: {, O+ G9 C/ w6 [

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板凳
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干细胞之家微信公众号
很好

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报纸
发表于 2011-3-8 20:30 |只看该作者
看那

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优秀会员

地板
发表于 2011-3-8 20:46 |只看该作者
这么给力啊 接力~

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发表于 2011-3-8 20:50 |只看该作者
怎么购买要八个包包 下载还要八个包包啊  物价上涨太快了

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asdfasfd
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发表于 2011-3-8 21:20 |只看该作者
非常3 J/ E5 P( Y3 J+ K2 z$ |
感谢楼主。

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就是看不了,可惜了。
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