TwoDimensionalGelElectrophoreticAnalysisfortheHumanPlasmaProteome
OverviewThis protocol is a detail description of the laboratory procedure in performing 2D gel electrophoresis for illustrating the protein profile of the human plasma proteome. Material1.Peripheral whole blood.2.Centrifuge machine3.Spectrophotometer4.Sample solubilization solution:(1).25.5 g urea + 50 ml distilled H2O(2).0.385 g DTT(3).2 ml NP-40(4).250 l carrier ampholyte (pH 3 10)(5).Trace amount Bromophen......阅读全文
Two－Dimensional－Gel－Electrophoretic－Analysis－for－the－Human－Plasma－Proteome
OverviewThis protocol is a detail description of the laboratory procedure in performing 2D gel electrophoresis for illustrating the protein profile of
2－Dimensional－Gel－Electrophoretic－Analysis－for－Chicken－Egg
Overview This protocol is a detail description of the procedure in performing 2D gel electrophoresis for illustrating the protein profile of the w
Two－dimensional－peptide－mapping
This specfic protocol is the latest incarnation of peptide mapping procedures that have been developed here in the TVL/MBVL of the Salk Institute over
Preparation－of－Stroma,－Thylakoid－Membrane,－and－Lumen－Fractions－from－...
Preparation of Stroma, Thylakoid Membrane, and Lumen Fractions from Arabidopsis thaliana Chloroplasts for Proteomic AnalysisFor many studies regarding
Twohybrid－analysis－of－genetic－regulatory－networks
1. Introduction and BackgroundThere is a great need for general methods to characterize the proteins that contemporary biology makes available. The li
Isolation－of－DL(Low－Density－Lipoprotein)－From－Human－Plasma
This procedure involves use of human plasma, a potentially dangerous source of blood-borne disease. Wear long-cuffed gloves, and eye-protection.Day on
二维聚丙烯酰胺凝胶电泳(twodimensional－polyacrylamide－gel－el
二维聚丙烯酰胺凝胶电泳技术结合了等电聚焦技术（根据蛋白质等电点进行分离）以及SDS－聚丙烯酰胺凝胶电泳技术（根据蛋白质的大小进行分离）。这两项技术结合形成的二维电泳是分离分析蛋白质最有效的一种电泳手段。通常第一维电泳是等电聚焦，在细管中（φ1～3 mm）中加入含有两性电解质、8M的脲以及非离子型去污
碳水化合物分析
Carbohydrate Assay (Hancock Laboratory) (Accessible only by IE)This protocol is used to determine the relative amounts of LPS CHO present in a given s
Sample－preparation－(analytical－gels)
Sample preparation and solubilization are crucial factors for the overall performance of the 2-D PAGE technique. Protein complexes and aggregates shou
QUALITATIVE－ANALYSIS－OF－DNA－FRAGMENTATION－BY－AGAROSE－GEL－ELECTROPHORESIS
1. IntroductionNuclear morphology changes characteristic of apoptosis appear within the cell together with a distinctive biochemical event: the endonu
双向电泳（twodimensional－electrophoresis）完整操作步骤
（一）第一向等电聚焦从冰箱中取-20℃冷冻保存的水化上样缓冲液（I）（不含DTT，不含Bio-Lyte）一小管（1ml/管），置室温溶解。2. 在小管中加入0.01g DTT， Bio-Lyte 4-6、5-7各2.5ml，充分混匀。3. 从小管中取出400ml水化上样缓冲液，加入100ml样品，充
Twohybrid－analysis－of－genetic－regulatory－networks2
2.2 Interaction mating - large scaleWith a few modifications, the procedure described above can be used to test for interactions between a single prey
Lipoprotein－Analysis－Week－2:－Electrophoresis
Lipoprotein Analysis Week 2: Electrophoresis IntroductionSDS polyacrylamide gel electrophoresis (SDS PAGE) will be used to assess the purification pr
Protocol－for－Protein－Extraction－for－proteomics
Protocol for Protein Extraction10 % w/v TCA/ acetone/ 0.07 % v/v -MercaptoethanolPlant cells are rich in compounds that interfere with the 2DE separat
RNA－analysis－on－nondenaturing－agarose－gel－electrophoresis
实验概要RNA analysis on non-denaturing agarose gel electrophoresis实验步骤1. The following gel electrophoresis conditions are recommended:- use 1X TAE buffer
RNA－analysis－on－nondenaturing－agarose－gel－electrophoresis
1. The following gel electrophoresis conditions are recommended:- use 1X TAE buffer instead of 1X TBE- use agarose gel in the concentration of 1.1%-1.
Comprehensive－identification－of－novel－proteins－and－Nglycosylation－sites八
References1. Fujita T, Kozuka-Hata H, Ao-Kondo H, Kunieda T, Oyama M, Kubo T: Proteomic analysis of the royal jelly and characterization of the fu
Peptide－map－prediction
Peptide map predictionIn identifying peptides from proteins with a known sequence, it is often useful to be able to predict how a peptide will migrate
Peptide－map－prediction
In identifying peptides from proteins with a known sequence, it is often useful to be able to predict how a peptide will migrate during electrophoresi
RNA－Electrophoresis
Electrophoresis through agarose or polyacrylamide gels is the standard way to separate, identify and purify nucleic acid fragments. The location of th
Study－on－a－TwoDimensional－Scanning－MicroMirror－and－Its－Application－...4
The target ranging is based on the phase-shift laser ranging method. The phase difference between the modulated signal and the reflected signal
双向电泳(twodimensional－electrophoresis,2DE)2
2．[操作步骤] 1. 将研磨管离心一分钟（不低于12000×g）弃上清。 2. 加入裂解液（200-300ul）充分vortex. 3. 再加入样品鼠脑组织（低于100mg）充分研磨。然后可以再加入裂解液至1ml.. 4. 将组织悬液离心5-10min(高于12000
Study－on－a－TwoDimensional－Scanning－MicroMirror－and－Its－Application－...2
In order to realize the decoupling measurement for two deflection angles and obtain the large piezoresistive coefficients for the high measureme
Study－on－a－TwoDimensional－Scanning－MicroMirror－and－Its－Application－...1
Study on a Two-Dimensional Scanning Micro-Mirror and Its Application in a MOEMS Target DetectorChi Zhang,* Zheng You, Hu Huang, and Guanhua LiAuthor i
Study－on－a－TwoDimensional－Scanning－MicroMirror－and－Its－Application－...3
2.4. CharacteristicsThe two resonance frequencies of the two-dimensional scanning micro-mirror are 216.8 Hz and 464.8 Hz, respectively, which are
双向电泳(twodimensional－electrophoresis,2DE)5
六、第二向 SDS-PAGE1．[基本原理]蛋白质在聚丙烯酰胺凝胶中电泳时，它的迁移率取决于它所带净电荷以及分子的大小和形状等因素。如果加入一种试剂消除电荷、形状等因素的影响，使电泳迁移率只取决于分子的大小，就可以用电泳技术测定蛋白质的分子量。1967年，Shapiro等发现在样品介质和聚丙烯酰胺凝
双向电泳(twodimensional－electrophoresis,2DE)6
我们本次实验使用的是银染。银染的方法种类很多，目前有文献报道的就有100多种。但是其准确的染色机制还不是特别的清楚。大致的原理是银离子在碱性pH环境下被还原成金属银，沉淀在蛋白质的表面上而显色。 由于银染的灵敏度很高，可染出胶上低于1 ng/蛋白质点，故广泛的用在2D凝胶分析上。待找到自己感
双向电泳(twodimensional－electrophoresis,2DE)1
一、蛋白质组学概论随着人类基因组计划的实施，生命科学步入了后基因组时代，出现了不同于以往经典生物实验科学的全新的研究方式─“生物大科学”。这种生物大科学的核心思想是整体性研究，即以生物体内某类物质为对象进行完整的研究。过去对生命活动的研究仅限于研究细胞内个别的基因或蛋白质，而基因组学和蛋白质组学的目
双向电泳(twodimensional－electrophoresis,2DE)3
2．[操作步骤]1. 将标准品BSA(5mg/ml)先稀释成0.5 mg/ml，2. 按0, 1, 2, 4, 8, 12, 16, 20μl分别加到96孔板中，加DDW补足到20μl。3. 加适当体积样品（4μl）到96孔板的样品孔中，加DDW到20μl。4. 各孔加入200μl G-250染色液
双向电泳(twodimensional－electrophoresis,2DE)4
由于合成载体两性电解质（synthetic carrier ampholyte SCA）是通过复杂的合成过程得到的，其重复性很难控制，由此不同批次之间会存在很大的变化，同一蛋白质在不同批 图-1. 等电聚焦的“聚焦效应” 次等电聚焦中所出现的位置有所偏差，这样作为双向电泳中的一向时就限