SizeandShapeofProteinMolecules4
Determining the Molecular Weight of a Protein Molecule—Combining S and R s à la Siegel and MonteWith the completion of multiple genomes and increasingly good annotation, the primary sequence of almost any protein can be found in the databases. The molecular weight of every protein subunit is therefore known from its sequence. But an experimental measure is still needed to determine if the nat......阅读全文
Bradford－Protein－Concentration－Assay
Bradford Protein Concentration Assayversion 01/07/2001Abbreviations:mcg = microgramsmcL = microlitersBSA = bovine serum albuminO.D. = optical densityd
Cyanogen－Bromide－digestion－of－protein
1. Proteins are TCA precipitated and washed with acetone, then dried.2. The CNBr should be brought to room temperature in the hood and used ONLY in th
Coupling－Antibodies－to－Protein－A－or－G
1. use 2 mg of antibody per ml wet beads (use appropriate antibody/protein A or G combination).2. mix antibodies with beads and bind at room temperatu
Protein－G－Purification－of－Antibodies
1. Reagent and Materials(1) Hi-Trap Protein G Column (Pharmacia Biotech #17-0404-01)(2) 20 mM Sodium Phosphate Buffer, pH 7.01.084 g NaH2PO4, anhydrou
Angiotensin－Protein－Kinase－Assay
James Hardwick's angiotensin assay protocolThis specific procedure was developed to assay the activity of the Lck kinase expressed from a retrovir
Basic－Protein－Chemistry－Techniques
实验概要Basic Protein Chemistry Techniques实验步骤Coomassie Blue Stain: (for gels) 1) Combine 225 ml Methanol with 225 ml ddH2O. 2) Add 0.5 grams of Coomassi
Mechanism－of－Protein－Import－into－the－Nucleus
Nuclear pore complexes (NPCs) are large proteinaceous assemblies that provide the only known portals for exchanging macromolecules between the nucleus
Protein－Expression－and－Purification－Protocol
Step 1: Transform appropriate DNA plasmid into BL21(DE3) E. coli cells. These cells must be competent. (Protocol for how to make competent cells.)a) T
In－Vitro－Protein－Ubiquitination－Assay
Ubiquitination is one of the most important posttranslational modifications in all eukaryote organisms. Ubiquitin-activating enzyme (E1), ubiquiti
Western－Blot－with－Platelet－Protein
OUTLINEWestern blot is a wide used technique to identify a target protein/s for the certain antibody.PROTOCOLPrepare platelets.Lyse washed platelets (
Protein－Immunolocalization－in－Maize－Tissues
The analysis of gene expression at transcript and at protein level is of outstanding importance in plant developmental biology. Proteins can be loc
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
Structure－of－Mitochondria
The cytoplasm of nearly all eukaryotic cells contain mitochondria, although there is at least one exception, the protist Chaos (Pelomyxa) carolinensis
Protein－detection－onto－PVDF－membranes
2-D PAGE and electroblotting onto PVDF membranes have become widely used techniques for the characterization of proteins. Recent improvements have all
COMPARISION－OF－DIFFERENT－PROTEIN－DETERMINATION－METHODS
COMPARISION OF DIFFERENT PROTEIN DETERMINATION METHODSCompanyMethodDetection RangeApplications -CompatibilityAssay protocolPrecautions-InterferencesA
Protein－Syntheses－in－Cell－Free－Systems
LEVEL IIIMaterialsSuspension culture of fibroblast cells (1 liter)35 mM Tris-HCl, pH 7.4, 140 mM NaCl (TBS buffer)10 mM Tris-HCl, pH 7.5, 10 mM KCl, a
Protein－concentration－of－Laemmli－gel－samples
Protein concentration of Laemmli gel samplesTo 10 µl boiled lysate (in Laemmli sample buffer) add 40µl water + 50µl 50% TCA. Ppt. 10 min. on ice. Spin
蛋白质（protein）概述
蛋白质是一种复杂的有机化合物，旧称“朊”。蛋白质这一概念最早是由瑞典化学家永斯·贝采利乌斯于1838年提出，但当时人们对于蛋白质在机体中的核心作用并不了解。1926年，詹姆斯·B·萨姆纳揭示尿素酶是蛋白质，首次证明了酶是蛋白质。第一个被测序的抗原肽蛋白质是胰岛素，由弗雷德里克·桑格完成，他也因此获得
Eccles:Protein－Lysates－from－Tissue
Cell Lysis Buffer5mL 0.1M Tris HCl pH 8 (10mM)0.44g NaCl (150mM)0.02g EDTA (1mM)0.5mL nonidet P40 (1% w/v)0.05g SDS (0.1% w/v)Make up to 50mL with MQH
Platelet－Amyloid－Precursor－Protein－Pathway
The amyloid -beta peptide (Ab), a proteolytic fragment of amyloid precursor protein (APP), is the major componenet of senile plaques, the hallmark of
Measurement－of－Green－Fluorescent－Protein－Expression
ReagentsCells to be studied expressing green fluorescent protein (GFP). Note that the same cell type without GFP is needed as control.Hoechst 33342 s
Mitochondria－Structure－and－Function－－Theoretical－Details
Mitochondria Structure and Function - Theoretical DetailsIn an attempt to be concise and understandable, introductory level courses and textbooks freq
线粒体的结构与功能
In an attempt to be concise and understandable, introductory level courses and textbooks frequently present concepts that are technically correc
troponin蛋白纯化－Protein－purification:－troponins
Overview TROPONINS The calcium-dependent regulatory protein complex located on the thin actin filaments of muscle comprises of TnC (17.8 kDa), TnI (2
UV－Absorbance－(280－nm)－－–－Protein－Determination
UV Absorbance (280 nm) – Protein Determination Simple and quick method to accurately quantitate total protein in purified material or approximately q
Protein－L树脂填料应用说明
Protein L也是一种免疫球蛋白结合蛋白，来源于从消化链球菌马格努斯。不同于Protein A和Protein G，Protein L结合的是抗体分子的轻链。由于没有重链的参与，比起Protein A和Protein G，Protein L可以结合更多种类型的抗体。Protein L可以
蛋白浓缩（protein－concentration）基本方法
蛋白浓缩方法基本有：丙酮沉淀法；免疫沉淀法；三氯醋酸沉淀法；硫酸铵沉淀法；（低温）有机溶剂沉淀法；聚乙二醇沉淀法；超滤法；透析法；离子交换层析和冷冻干燥法…… 1.丙酮沉淀法；三氯醋酸沉淀法 试验要求的仪器简单，但是常常导致蛋白质变性。 2.免疫沉淀法：得有特异性抗体！ 3.硫酸铵沉淀法
Protein－extraction－from－whole－tissues－for－IEF
Modified from that of Jay Thelen - University of Missouri-ColumbiaPhenol extraction followed by methanolic ammonium acetate precipitation - an effecti
Total－Protein－Extraction－with－TCAAcetone
We describe a procedure allowing extraction of total proteins that performs efficiently with a large variety of plant tissues, based on simultaneo
Purification－of－Antiserum－or－Ascites－by－Protein－A/G－Chromatography
1、Required Materials and Equipment(1) Protein A or G agarose gel column (10 ml or 5 ml of packed beads; see guidelines below for choice of protein A o