蛋白质提取与制备(ProteinExtractionandPreparation)3
水溶液提取：大部分蛋白质均溶于水、稀盐、稀碱或稀酸溶液中。因此蛋白质的提取一般以水为主。稀盐溶液和缓冲溶液对蛋白质稳定性好、溶度大，也是提取蛋白质的最常用溶剂。盐溶液提取：以盐溶液及缓冲液提取蛋白质进常注意下面几个因素。盐浓度等渗盐溶液尤以0.02～0.05mol/L 磷酸盐缓冲液和碳酸盐缓冲液常用。0.15mol/L 氯化钠溶液应用也较多。如6-磷酸葡萄糖脱氢酶用0.1mol/L 碳酸氢钠液提取等。有时为了螯合某些金属离子和解离酶分子与其他杂质的静电结合，也常使用枸橼酸钠缓冲液和焦磷酸钠缓冲液。有些蛋白质在低盐浓度下浓度低，如脱氧核糖核蛋白质需用1mol/L 以上氯化钠液提取。总之，只要能溶解在水溶液中而与细胞颗粒结合不太紧密的蛋白质和酶，细胞破碎后选择适当的盐浓度及PH，一般是不难提取的。只有某些与细胞颗粒上的脂类物质结合较紧的，需采用有机溶剂或加入表面活性剂处理等方法提取。PH 值：蛋白质提取液的PH ......阅读全文
细胞膜蛋白质提取方法
NRC Institute for Biological SciencesTriton X-114 extraction protocol (Hydrophobic protein preparation)Ressuspend cells in Solution A (dil 1/8) and ad
组织学——显微解剖
Laser Capture Microdissection (LCM)Introduction to LCM (BJMU) Preparation, LCM and RNA/DNA extraction of Frozen Tissue Sections (NIH Laser Capture M
Isolation－of－Nonmuscle－Actin
General Preparation1. Prepare buffers and have them cold. Make 50X stock of Buffer G and dilute as needed from frozen aliquots of 50X Buffer G.1X Extr
细胞组分和细胞器——染色体
Chromosomal DNA Prep : cultured cells/tissue samples (Mike A Dyer)This protocol was developed for cultured cells but should be appropriate for dissoci
表达蛋白（Expressed－protein）的分离与纯化
大肠杆菌表达蛋白以可溶和不溶两种形式存在，需要不同的纯化策略。现在，许多蛋白质正在被发现而事先并不知道它们的功能，这些自然需要将蛋白质分离出来后，进行进一步的研究来获得。分析蛋白质的方法学现已极大的简化和改进。必须承认，蛋白质纯化比起DNA 克隆和操作来是更具有艺术性的，尽管DNA 序列具有异乎
蛋白质二级结构预测(protein－secondary－structure－prediction)
蛋白质二级结构的预测开始于20世纪60年代中期。二级结构预测的方法大体分为三代，第一代是基于单个氨基酸残基统计分析，从有限的数据集中提取各种残基形成特定二级结构的倾向，以此作为二级结构预测的依据。第二代预测方法是基于氨基酸片段的统计分析，使用大量的数据作为统计基础，统计的对象不再是单个氨基酸残基，而
蛋白质二级结构(protein－secondary－structure)预测软件
蛋白质二级结构的预测通常被认为是蛋白结构预测的第一步，二级结构是指α螺旋和β折叠等规则的蛋白质局部结构元件。不同的氨基酸残基对于形成不同的二级结构元件具有不同的倾向性。按蛋白质中二级结构的成分可以把球形蛋白分为全α蛋白、全β蛋白、α＋β蛋白和α/β蛋白等四个折叠类型。预测蛋白质二级结构的算法大多以已
组织学——组织制备
· Histological techniques (William H. Heidcamp)Very detailed guide to histological techniques, like fixation, dehydration, embedment and subs
Neuroregulin－receptor－degredation－protein1－Controls－ErbB3－receptor
The neuregulins comprise a subfamily of at least four epidermal growth factor (EGF)-like growth factors that influence a variety of cellular events, i
蛋白质、多肽提取分离
1 分离方法 采取何种分离纯化方法要由所提取的组织材料、所要提取物质的性质决定。对蛋白质、多肽提取分离常用的方法包括：盐析法、超滤法、凝胶过滤法、等电点沉淀法、离子交换层析、亲和层析、吸附层析、逆流分溶、酶解法等。这些方法常常组合到一起对特定的物质进行分离纯化，同时上述这些方
蛋白质、多肽提取分离
1 分离方法 采取何种分离纯化方法要由所提取的组织材料、所要提取物质的性质决定。对蛋白质、多肽提取分离常用的方法包括：盐析法、超滤法、凝胶过滤法、等电点沉淀法、离子交换层析、亲和层析、吸附层析、逆流分溶、酶解法等。这些方法常常组合到一起对特定的物质进行分离纯化，同时上述这些方法也是蛋白、多肽类物
Sumo化蛋白定量试剂盒—小泛素化研究
众所周知，泛素（ubiquitin, Ub）是一类高度保守的小蛋白, 可与靶蛋白的赖氨酸残基共价连接, 形成多聚泛素链行使指导蛋白质降解的功能。类似于泛素化修饰过程, 小泛素相关修饰物(small ubiquitin related modifier, SUMO）也可以共价修饰靶蛋白的赖
Streamlined－DNA－Extraction－Protocol
This method is derived from a procedure developed by Toby Bradshaw and the Poplar Molecular Genetics Cooperative. We have tested the procedure wit
Fungal－Genomic－DNA－Extraction
OverviewHigh throughput of many fungal isolates can be achieved by growing axenic cultures in either (a) 1.5mL microfuge tubes, half full with liquid
Fungal－Genomic－DNA－Extraction
OverviewHigh throughput of many fungal isolates can be achieved by growing axenic cultures in either (a) 1.5mL microfuge tubes, half full with liquid
DNA－Extraction－from－Tissue
实验概要DNA extraction from tissue.主要试剂Extraction buffer100 mM Tris-HCl (pH 8.0) 100 mM EDTA (pH 8.0) 100 mM Na-Phosphate (pH 8.0) 1.5 M NaCl1% CTAB
Fungal－Genomic－DNA－Extraction
实验概要This procedure does not require phenol extraction. The DNA is pure enough for restriction digests, PCR and genomic library construction.High t
Arabidopsis－RNA－extraction－protocol
1-2 g fresh material, freezer-dried, ground with 0.2g sand (if necessary), and then homogenized with 10ml RNA extraction buffer (see below). Spin
Automated－Genomic－DNA－Extraction
实验概要This section provides a general protocol for automated isolation of genomic DNA from 10-20 µl blood samples in a 96-well format using the Charge
Genomic－DNA－Extraction－－PureLink™
实验概要The PureLink™ Genomic DNA Purification Kit allows rapid and efficient purification of genomic DNA. The kit is designed to efficiently isolate g
DNA－EXTRACTION－PROCEDURE－－GENERAL
Grow cells overnight in 500 ml broth medium.Pellet cells by centrifugation, and resuspend in 5 ml 50 mM Tris (pH 8.0), 50 mM EDTA.Freeze cell suspensi
DNA－Extraction－from－Blood
实验概要The ChargeSwitch® gDNA Purification Kits allow rapid and efficient purification of genomic DNA from small volumes of human blood. After preparin
Arabidopsis－RNA－extraction－protocol
1-2 g fresh material, freezer-dried, ground with 0.2g sand (if necessary), and then homogenized with 10ml RNA extraction buffer (see below).Spin at 8,
蛋白质的蛋白质的提取技术
选择材料及预处理 以蛋白质和结构与功能为基础，从分子水平上认识生命现象，已经成为现代生物学发展的主要方向，研究蛋白质，首先要得到高度纯化并具有生物活性的目的物质。蛋白质的制备工作涉及物理、化学和生物等各方面知识，但基本原理不外乎两方面。一是得用混合物中几个组分分配率的差别，把它们分配到可用机械方
French－Pressure－Cell超高压细胞破碎仪应用
一、细胞破碎应用 Cell Rupture, Disruption, Lysis are methods for disrupting cell walls and cell membranes to release biological molecules, whilst maintainin
Preparation－of－human－platelets
Preparation of human platelets 1. Human blood was taken from drug-free volunteers on the day of the experiment using acidic citrate dextrose
HELPER－PHAGE－PREPARATION
HELPER PHAGE PREPARATION1. Grow an overnight of NM522 in NZCYM medium.2. Dilute overnight 1:100 and grow to an A600 = 0.3 (@2.5 x 108 cells/ml).3. Inf
Lambda－DNA－Preparation
Lambda DNA PreparationThis is a plate method that gives very good yield for cloning. I have combined the Promega and Maniatis protocols.Solutions T-TY
Preparation－of－Polyacrylamide－Gels
1. Prepare 20X TBE as:216 g Tris Base110 g Boric Acid80 mL 500 mM EDTA, pH 8.0700 mL ddH2OMix. Bring volume to 1 L. Autoclave.2. Prepare Acrylamide so
Competent－Cell－Preparation
实验概要Competent cells are those that possess more easily altered cell walls that DNA can be passed through easily. These cells readily incorporate f