SizeandShapeofProteinMolecules2
The sedimentation coefficient of a protein is a measure of how fast it moves through the gradient. Increasing the mass of the protein will increase its sedimentation, while increasing its size or asymmetry will decrease its sedimentation. The relationship of S to size and shape of the protein is given by the Svedberg formula:(4.1)M is the mass of the protein molecule in Dalton; N o is Av......閱讀全文
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
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
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
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
Basic－Protein－Chemistry－Techniques
Coomassie Blue Stain:? (for gels)?1) Combine 225 ml Methanol with 225 ml ddH2O.?2) Add 0.5 grams of Coomassie Blue.?3) Just before use, add 50 ml acet
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 (
Biorad－Protein－Assay:－Bradford
Biorad Protein Assay: BradfordStandards: 1 mg/ml BSA stock- dilute 1:10 to get 0.1 mg/ml BSAAdd To get H-2O20 μl 2 μg/ml 780 μl40 μl 4 μg/ml 760 μl60
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
RNA－Electrophoresis
Electrophoresis through agarose or polyacrylamide gels is the standard way to separate, identify and purify nucleic acid fragments. The location of th
Immunoblotting－(Western－Blotting)
實驗概要We provide a protocol for SDS-PAGE, Protein Blotting, Immuno-Detection.主要試劑1.?0.3 M TRIZMA? base (Product No. T1503), 20% methanol.2.?0.025 M TRIZ
Activation－and－Expansion－of－Human－Treg－Cells
實驗概要This protocol is intended for activation and expansion of human Treg cells isolated with the Dynal? CD4 CD25 Treg Kit (Cat. no. 113.23D). The exp
Genomic－Libraries
Genomic DNA libraries?Size of some genomes and chromosomes:Comparative Sequence Sizes(Bases)(yeast chromosome 3)350 ThousandEscherichia coli (bacteriu
Gel－Electrophoresis－of－DNA
What is Electrophoresis?Electrophoresis is a technique used in the laboratory that results in the separation of charged molecules. In this CyberLab we
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
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
蛋白質（protein）概述
蛋白質是一種復雜的有機化合物，舊稱“朊”。蛋白質這一概念最早是由瑞典化學家永斯·貝采利烏斯于1838年提出，但當時人們對于蛋白質在機體中的核心作用并不了解。1926年，詹姆斯·B·薩姆納揭示尿素酶是蛋白質，首次證明了酶是蛋白質。第一個被測序的抗原肽蛋白質是胰島素，由弗雷德里克·桑格完成，他也因此獲得
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
COMPARISION－OF－DIFFERENT－PROTEIN－DETERMINATION－METHODS
COMPARISION OF DIFFERENT PROTEIN DETERMINATION METHODSCompanyMethodDetection??RangeApplications?-CompatibilityAssay protocolPrecautions-InterferencesA
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
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
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
重組DNA的分離、克隆與測序實驗手冊2
C. Restriction digestionRestriction enzyme digestions are performed by incubating double-stranded DNA molecules with an appropriate amount of restrict
Immunostaining－Thin－Layer－Chromatograms－Of－Glycolipids
Immunostaining Thin Layer Chromatograms Of GlycolipidsJohn L. Magnani~GlycoTech Corporation, Rockville, Maryland 20850Immunostaining thin layer chroma
Dynabeads?－Streptavidin－Trial－Kit（二）
3. General Immobilization Protocol??????????? Wash the Dynabeads? before use.??????????????????a)????? Add the biotinylated molecule to the washed Dyn
Activation－of－cAMPdependent－protein－kinase,－PKA
G-protein coupled receptors (GPCRs) are one of the largest gene families of signaling proteins. Residing in the plasma membrane with seven transmembra
Basic－procedures－for－bacteria－culture1
A. Phenol extraction of DNA samplesPhenol extraction is a common technique used to purify a DNA sample (1). Typically, an equal volume of TE-saturated
Molecules：阿司巴汀緩解多柔比星心臟毒性
多柔比星（Dox）是用于治療各種癌癥的有效化學藥物。但是，由于潛在的致命心臟毒副作用，其臨床應用常常受到限制。越來越多的證據表明，腫瘤蛋白p53（p53），腺苷單磷酸激活蛋白激酶（AMPK），核蛋白p62（p62）和雷帕霉素哺乳動物靶標（mTOR）是Dox誘導細胞凋亡的關鍵介質，并參與隨后自噬的
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
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
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