新型纳米金蛋白质定位微试管芯片和计算与信号传导

内容简介

《新型纳米金蛋白质定位微试管芯片和计算与信号转导》作者原创一种新型纳米金微芯片，并整合细胞核和细胞质蛋白的纯化结果和生物计算技术（如DB索引算法，由距离矩阵、色彩相似性等决定聚类边界），通过分子标注和击打次数，建立细胞信号转导模型。该系统具有高特异性、高敏感性、高重复性、高稳定性、高信号性、样本检测量大、简便性、安全性、多参数实时定量的特点，通过计算后可建立分子机制网络。

目录

Chapter 1 Introduction
1.1 Antibody Array Technologies and Applications
1.2 Protein Phosphorylation, Localization and Function
1.3 Interaction Proteomics and Pathway Building
1.3.1 BMP2 Signaling
1.3.2 The Effect of STI571 on Cell Signaling

Chapter 2 Technologies
2.1 Materials
2.1.1 Cell Lines
2.1.2 Chemicals
2.1.3 Antibodies
2.1.4 Protein-Molecular Weight Marker
2.1.5 Apparatus
2.2 Methods
2.2.1 Cell Culture
2.2.2 Cell Numbers and Experimental Format
2.2.3 Cytoplasmic and Nuclear Protein Extracts
2.2.4 Determination of Protein Concentration
2.2.5 Ponceau S Staining
2.2.6 Western Analysis
2.2.7 Immunoprecipitation
2.2.8 Cell Cycle Analysis
2.3 Silver Staining in ArrayTube
2.3.1 Biotinylation of Cellular Proteins
2.3.2 Incubation of Protein Extracts
2.3.3 Silver Staining
2.3.4 Statistical Analysis

Chapter 3 New Cytoplasmic and Nuclear BMP2-Signaling Pattern in U937 Cells
3.1 Modulation of Phosphorylation and Localization in BMP2-Treated U937 Cells
3.1.1 Total p-Tyrosine and p-p38, p38 Increased: Increasing Cytoplasm and Nucleus
3.1.2 Total p-ERK and p-JNK Increased: Increasing Cytoplasm and Unchanged Nucleus
3.1.3 Total p-Akt Increased: Increasing Cytoplasm and Decreasing Nucleus
3,1.4 Total p-Smadl, p-Smad2/3, c-Myc and p-STAT3 Unchanged:Decreasing Cytoplasm and Increasing Nucleus
3.1.5 Total p-p70S6 Unchanged: Increasing Cytoplasm and Decreasing Nucleus
3.2 BMP2 Signaling in U937 Cells
3.2.1 BMP2 Activates Akt, ERK and INK Pathway in U937 Cells
3.2.2 BMP2 Activates Smadl and Smad2/3 Networks, p38 Network and c-Myc, Tyrosine Network in U937 Cells
3.2.3 BMP2 Inhibits p70S6 Signaling in U937 cells
3.3 BMP2 Signaling Model in U937 Cells

Chapter 4 New Cytoplasmic and Nuclear BMP2-Signaling in MCF7 Cells
4.1 Modulation of Phosphorylation and Localization in BMP2-Treated MCF7 Cells
4.1.1 Total p-JNK, p-Smadl, p-Smad2/3, p-p38 and p38 Unchanged:Increasing Cytoplasm and Decreasing Nucleus
4.1.2 p-Tyrosinc, p-STAT3, p-ERK and p-p70S6 Increased:Increasing Cytoplasm and Unchanged Nucleus
4.1.3 Total c-Myc Unchanged: Slightly Increasing Nucleus and Slightly Decreasing Cytoplasm
4.1.4 Total p-Akt Unchanged: Unchanged Nucleus and Cytoplasm
4.2 BMP2 Signaling in MCF7 Cells
4.2.1 BMP2 Activates p70S6 and ERK Signaling, Tyrosine and STAT3 Network in MCF7 Cells ..
4.2.2 BMP2 Inhibits p38, JNK and Smad Signaling in MCF7 Cells
4.2.3 BMP2 Activates c-Myc Signaling in MCF7 Cells
4.3 BMP2 Signaling Model in MCF7 Cells

Chapter 5 New Cytoplasmic and Nuclear ST1571 Signaling in K562 Cells
5.1 Modulation of Phosphorylation and Localization in STI571-Treated K562 Cells
5.1.1 Total p-Akt and p-p70S6 Decreased: Decreasing Cytoplasm and Nucleus
5.1.2 Total p-p38, p38, c-Myc, p-Tyrosine, p-Smadl and p-Smad