BacterialExpressionofIRS1containingGSTfusionProteins
1. Grow cells in 5ml (or more) of LB-Amp overnight for a starter culture. 2. Grow larger culture using the overnight culture as a seeding culture. The culture is grown in LB-Amp medium. Aerate well with shaking at 37 C until OD at 600nm is ~0.6. 3. Add the appropriate amount of IPTG stock solution to the culture to obtain a final IPTG concentration of ~2mM. 4. Continue sha......阅读全文
Bacterial－Expression－of－IRS1－containing－GSTfusion－Proteins
1. Grow cells in 5ml (or more) of LB-Amp overnight for a starter culture. 2. Grow larger culture using the overnight culture as a seeding culture.
Bacterial－Expression－of－GSTfusion－Proteins
1. Grow cells in 5ml (or more) of LB-Amp overnight for a starter culture. 2. Grow larger culture (100x volume of starter culture) using the overnigh
Preparation－of－Bacterial－Proteins－for－Analysis－by－2DPAGE
The following protocol has been developed for preparing soluble bacterial proteins in a form suitable for analysis by 2D-PAGE. The procedure was princ
Expression－Library－Screening－(Procaryotic)－Using－APFusion－Proteins
Outline:Bacteriophage lambda is a linear double stranded DNA, approximately 50 kB in size. The two sticky ends help the phage to recircularize after e
GST融合蛋白的准备
Preparation of Glutathione-S-Transferase (GST) Fusion ProteinsMargret B. Einarson and Elena N. Pugacheva Foxx Chase Cancer Center, Philadelphia, PA 19
亲和层析实验技术方法
INTRODUCTIONThis protocol describes a method for removing antibodies that react with bacterially encoded proteins by passing a crude preparation of im
GST融合蛋白纯化——筛选表达株
Purification of GST fusion proteins in E.coli GSTSugden lab,McArdle Laboratory for Cancer Research ,University of Wisconsin-Madison Medical SchoolScre
蛋白质相互作用
Interaction Trap/Trap Two-Hybrid System· Yeast Two-Hybrid System (Finley Lab)This is one of the most comprehensive and detailed guide to yeast
Bacterial－transformation
IntroductionTransformation is the process of introducing foreign DNA (e.g plasmids, BAC) into a bacterium. Bacterial cells into which foreign DNA can
Analysis－of－Proteins－using－Small－Format－2D－Gel－Electrophoresis
Preparation of protein samplesIntracellular virus proteinsThe following method has been developed principally for the analysis of intracellular protei
Biosynthesis－of－Cysteine－from－serine－in－bacteria－and－plants
In animals cysteine is synthesized from homocysteine, a produce of the essential amino acid methionine. In the absence of dietary methionine, animals
Thrombin－Cleavage－of－GSTFusion－protein
INTRODUCTIONIn many cases the cleavage can be performed using the free intact fusion, or in same cases with the fusion protein bound to a matrix. The
蛋白质提取和纯化
蛋白质提取和纯化（主要内容如下）Protein Extraction Protein PurificationProtein PrecipitationColumn PreparatioinQ & A Posted in the Method ForumProtein ExtractionWhole
HypoxiaInducible－Factor－in－the－Cardiovascular－System
Hypoxia (or low O2 levels) affects various pathologies. First, tissue ischemia, a variation in O2 tension caused by hypoxia/reoxygenation, can lead to
Bacterial－glycerol－stocks
To 2mls of mid-log culture or 1ml of freshly saturated culture add 1 ml(or an equal volume) of glycerol solution, mix gently, then freeze rapidly in l
Bacterial－Colony－PCR
Bacterial Colony PCRObjective:This protocol allows rapid detection of transformation success when primers are available to allow determination of corr
Bacterial－cell－culture
MaterialsGlass culture tubes with metal caps and labelsGrowth medium, from media room or customizedGlass pipette tubesParafilmEquipmentVortexerFireboy
Streaking－Bacterial－Stocks
Principle:Bacterial cells are streaked onto a medium to obtain an independent isolate. This is done to reduce the likelihood of working with a culture
Purification－of－MBP－(maLTosebinding－proteins)－Fused－Proteins
Express fusion proteins as per the GST-fused protocol up to Step 7 (Day 3). All steps in protein purification should be done at 4° C unless otherwise
全细胞靶点筛选抗生素新药的方法
A target-specific whole cell assay for antibacterial drug discoveryLorraine HernandezSrinivas KodaliDoris CullySheo SinghJun Wang , jun_wang2@merck.co
Expression－L19－using－Pichia－pastoris
Pichia pastoris is a methylotropic yeast used to express high amounts of protein. Secreted expression is achieved with a cleavalable faktor. To yield
Purification－of－Kar3－Motor－Domain－Protein
Purification of Kar3 Motor Domain ProteinMaterialsInduced cells (2 - 5 g pellet of pET/Kar3 in BL31(DE3)pLysS host cells) (See note #1)HEM buffer =10
Bacterial－Media－Solutions－and－Stocks
3 agar (200 ml)Add 6 grams agar to 200 ml deionized water. Autoclave to sterilize.1.6 agar (200 ml)Add 3.2 grams agar to 200 ml deionized water.Autocl
Preparing－Overnight－Bacterial－Culture
Materials:Sterile LB medium (Luria-Bertani Medium) with or without antibiotic:water 500 mlbacto-tryptone 10 gbacto-yeast extracts 5 gsodium chloride 1
Production－of－Antibody－Fragments－in－Arabidopsis－Seeds
Plants offer a number of attractive benefits over conventional mammalian or bacterial cell culture systems for the production of valuable pharmace
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
MSI1基因编码功能及结构描述
这个基因编码一个含有两个保守的串联rna识别基序的蛋白质。其他物种中的类似蛋白作为rna结合蛋白发挥作用，在转录后基因调控中发挥中心作用。该基因的表达与胶质瘤和黑色素瘤的恶性程度和增殖活性有关。该基因的假基因位于染色体11q13上。[由RefSeq提供，2008年7月]This gene encod
MSI1基因突变与药物因子介绍
这个基因编码一个含有两个保守的串联rna识别基序的蛋白质。其他物种中的类似蛋白作为rna结合蛋白发挥作用，在转录后基因调控中发挥中心作用。该基因的表达与胶质瘤和黑色素瘤的恶性程度和增殖活性有关。该基因的假基因位于染色体11q13上。[由RefSeq提供，2008年7月]This gene encod
Small－Leucinerich－Proteoglycan－(SLRP)－molecules
The small leucine-rich proteoglycans (SLRPs) are a family of proteins that are present in extracellular matrix and that share in common multiple repea
Long－Term－Storage－of－Bacterial－Strains
Purpose:Bacterial strains may be stored indefinitely at low temperatures (- 20 degrees C and -80 degrees C) in 15 to 40 glycerol. It is lab policy to