PurificationofMBP(maLTosebindingproteins)FusedProteins
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 stated.Pre-wash 10-20 ml of New England Biolabs amylose resin with 20 mM Tris pH7.4, 200 mM NaCl, 10 mM b ME, 1mM EDTA.Pellet the cellular debris (SS34, 13000 rpm, 15’) from the lysate (Step 7).Dilute the supernatant to 100 ml with 20mM Tris pH7.4, 200 mM NaCl......阅读全文
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
Purification－of－GST－Fused－Proteins
Day 1Set up an overnight culture in 100 ml LMM broth or 100 ml terrific broth containing 100ul 100 mg/mlAmpDay 2Add 40-50 ml o/n culture to 1 lt terri
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
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
酵母双杂交系统
· Yeast Two-Hybrid System (Finley Lab)This is one of the most comprehensive and detailed guide to yeast two-hybrid system technique with intro
蛋白表达
Protein Construct Expression and Purification Procedures (Gimila's Lab) Protein Expression (Mark's Lab) · Purification of GST Fused
Production－of－Recombinant－Proteins－in－SuspensionCultured－Plant－Cells
Plants have emerged in the past decade as a suitable alternative to the current production systems for recombinant pharmaceutical proteins and, toda
Antibody－Purification
This protocol includes an ammonium sulfate cut, affigel blue chromatography and affinity chromatography.1. Solutions(1) Affigel Blue Prewash0.1 M acet
Protein－A－Purification－of－Antibody
1. Reagents(1) Affi-gel Protein-A Agarose (BioRad #153-6153)(2) MAPS II Binding Buffer (BioRad # 153-6161)(3) 0.314 g/ml diH2O(4) MAPS II Elution Buff
Protein－purification;－actin
Protein purification; actin Overview ACTINThe most abundant muscle and non-muscle cytoskeletal protein. MW 42 kDa, 374/375 amino acids; various
Purification－of－mAb－(IgG)
1. Materials(1) Antibody 7E3, 2L sup grown in flasks, frozen and thawed overnight.(2) BioRad Affi-Gel Protein A MAPS II Buffers cat. #1530-6160 ($161.
Purification－of－Demethylated－Sphingomyelin
I. Lower, chloroform phase:1) Dry on rotovapor system with house vacuum lines. It is not necessary to dry sample completely, but sufficiently to yield
Synaptic－Proteins－at－the－Synaptic－Junction
The postsynaptic density (PSD) is a submembranous structure at the postsynaptic membrane mainly at the excitatory synapses. The neurotransmitter recep
Column－Purification－of－Demethylated－Sphingomyelin
Packing column:1) To 20 g of 100-200 mesh Bio-Sil A silica gel add 80 mls of chloroform.2) Place a small portion of glass wool at the base of the colu
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
Antigen－Affinity－Purification－of－Antibodies
实验概要To acquire purified antibodies (This method typically yields >95% pure specific antibodies ).实验原理 Cytokines are signaling proteins necessary for
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
Factor－Xa－Cleavage－of－MBPFusion－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
如何选择合适的MBP标签抗体？
大肠杆菌麦芽糖结合蛋白（ Maltose Binding Protein，MBP）来源于大肠杆菌malE基因编码的蛋白，是细菌麦芽糖转运系统的成员之一，主要负责麦芽糖的摄取及分解代谢。malE基因产物能与多种蛋白融合，是进行表达研究的有效媒介，由于MBP融合蛋白原核表达载体具有表达效率高，易于纯
DNA－Immunoprecipitation－for－the－Determination－of－DNABinding－Specificity
Andrea J. Gossett and Jason D. Lieb1Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA1Corresponding autho
Phosphoproteins－pr...
实验概要The following procedure provides a method of detection of phosphorylated proteins.实验步骤1. To a sample of protein solution containing 1-100 ng of
Crystallization－of－Kinesin－Family－Motor－Proteins
Motor proteins of several kinesin family groups have now been crystallized: monomeric Kinesin-1 motor domains from human, rat andNeurospora (Kull et a
Acetylation－(or－Succinylation)－of－Amino－Groups－on－Proteins
Acetylation (or Succinylation) of Amino Groups on ProteinsREFERENCE: Hanock and Benz. 1986. BBA. 860:699-707.PURPOSE: Derivitization of amino groups t
大鼠髓鞘碱性蛋白(MBP)ELISA检测法
大鼠髓鞘碱性蛋白(MBP)ELISA试剂盒 (用于血清、血浆、细胞培养上清液和生物体液内) 原理本实验采用双抗体夹心 ABC-ELISA法。用抗大鼠 MBP 单抗包被于酶标板上，标准品和样品中的 MBP与单抗结合，加入生物素化的抗大鼠MBP，形成免疫复合物连接在板上，辣根过氧化物酶标记的Strept
DNA－Purification－from－Agarose－Gels
1. Separate DNA fragments in an agarose gel cast with 0.5 mg/mL Ethidium bromide. Locate bands with a hand-held long-wave UV lamp.2. Slice the gel wit
Purification－of－human－mononuclear－cells－and－neutrophils
PurposeMaterials10ml 6% dextran + 7ml citrate/citric acidDextran: T500 --> 6g+100ml PBSCitrate solution: 25g Na Citrate + 8g citric acid + 500 ml PBS4
CD2的纯化－Purification－of－CD2
Purification of CD21) Cells are pelleted (~3 - 4000 g for 10 min.) and resuspended in pre-chilled 50 mM malonate (pH 5.2-5.3) with 1mM EDTA (~40ml of
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
BTG－family－proteins－and－cell－cycle－regulation
BTG2 is found to be one of the immediate early genes up-regulated by neural growth factor (NGF) and epidermal growth factor (EGF). Its transcriptional
猪髓鞘碱性蛋白(MBP)ELISA试剂盒
猪髓鞘碱性蛋白(MBP)ELISA试剂盒 (用于血清、血浆、细胞培养上清液和生物体液内) 原理本实验采用双抗体夹心 ABC-ELISA法。用抗猪 MBP 单抗包被于酶标板上，标准品和样品中的 MBP与单抗结合，加入生物素化的抗猪MBP，形成免疫复合物连接在板上，辣根过氧化物酶标记的Streptavi