ThrombinCleavageofGSTFusionprotein
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 amount of thrombin, temperature and length of incubation must be calibrated for each system. Samples must be removed at various time points and analyzed by PAGE-SDS to estimate the yield, purity and extent of thrombin digestion. Excess thrombin may result in unwant......阅读全文
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
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
Fibrinolysis－Pathway
Clot formation and fibrinolysis is a balance of plasmin activation/inhibition and thrombin-thrombomodulin activity that regulates fibrin polymer forma
GSP融合蛋白的准备
GST Fusion Protein PrepItalics indicate optional steps especially useful for the analysis of untested proteins.GROWTH AND HARVESTING OF BACTERIAAdd 10
Transcriptional－activation－of－dbpb－from－mRNA
Endothelial cells respond to treatment with the protease thrombin with increased secretion of the PDGF B-chain. This activation occurs at the transcri
分子克隆蛋白表达实验指南（十二）
– Note: The yield of fusion protein can be estimated by measuring the absorbance at 280 nm. The GST affinity tag can be approximated by 1 A280 Å 0.5
分子克隆蛋白表达实验指南（十一）
SDS-PAGE胶样品排列： MarkerUII 37CUI’I’ Marker：低分子量蛋白marker，上样10ul UI：未诱导菌液，上样10ul。任取37C和20C中一个 I：诱导后对照，上样10ul UI’, I’代表G
Extrinsic－Prothrombin－Activation－Pathway
Blood coagulation or clotting takes place in 3 essential phases. The first phase is the activation of a prothrombin activator complex. The second phas
Intrinsic－Prothrombin－Activation－Pathway
Blood coagulation or clotting takes place in 3 essential phases. The first phase is the activation of a prothrombin activator complex. The second phas
Thrombin－signaling－and－proteaseactivated－receptors
Thrombin is an extracellular protease that is involved in the clotting of blood and inflammation through its action on platelets and endothelial cells
James－Hardwick－CNBr－Cleavage－Procedure
1. Immunoprecipitate the protein and run it on a preparative gel. CNBr cleavage must be done with protein transferred to a nitrocellulose filter. Neit
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
Antibody－Purification－using－Protein－A,－Protein－G,－or－Protein－L－Agarose
实验概要This protocol is designed as a quick purification method for antibodies from mammalian sera, ascites, and cell culture supernatants. It should
Antibody－Purification－using－Protein－A,－Protein－G,－or－Protein－L－Agarose
实验概要This protocol is designed as a quick purification method for antibodies from mammalian sera, ascites, and cell culture supernatants主要试剂 Protein
人凝血酶(Thrombin)ELISA试剂盒
人凝血酶(Thrombin)ELISA试剂盒 (用于血清、血浆、细胞培养上清液和其它生物体液内) 原理本实验采用双抗体夹心 ABC-ELISA法。用抗人Thrombin 单抗包被于酶标板上，标准品和样品中的 Thrombin与单抗结合，加入生物素化的抗人Thrombin，形成免疫复合物连接在板上，辣
Radioiodination－of－protein
Radioiodination (by Jun Takagi,6/16/2000)Purpose and backgroundsPrinciple of radioiodinationAddition of oxidizing reagents (such as chloramine-T or pe
Protein－Electrophoresis
DefinitionAmino acids, nucleotides, polypeptides, and other compounds in a colloidal state can be separated by the application of external voltages wh
Protein－Crystallization
Background:Proteins, like many molecules, can be prompted to form crystals when placed in the appropriate conditions. In order to crystallize a protei
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.
The－Determination－of－Proteinprotein－Interactions－by－the－Matingbased－...
Dynamic and reversible protein–protein interactions have a pivotal function in all living cells. For instance, protein–protein interactions are in
人凝血酶（Thrombin）酶联免疫分析（ELISA）
人凝血酶（Thrombin）酶联免疫分析（ELISA）试剂盒使用说明书本试剂仅供研究使用 目的：本试剂盒用于测定人血清，组织及相关液体样本中凝血酶（Thrombin）的含量。实验原理： 本试剂盒应用双抗体夹心法测定标本中人凝血酶（Thrombin）水平。用纯化的人凝血酶（Thromb
Actin－Capture－Assay
David AmbergDialyze purified GST fusion proteins and actin into PBS + 1mM MgCl2 .Mix 5ug actin into 50ul total volume binding buffer.Mix 5ug GST-fusio
凝血酶切割位点的定义
中文名称凝血酶切割位点英文名称thrombin cleavage site定 义基因工程表达系统中融合蛋白常用的连接序列，凝血酶处理可将融合蛋白中目的蛋白与非目的蛋白的肽段分离，最适切割位点是：P4-P3-P2-Arg↓-P1′-P2′，式中P4和P3为疏水氨基酸，P1′和P2′为非酸性氨基酸，↓
Eukaryotic－protein－translation
The scanning translation initiation model suggests that 40S ribosomal subunit preloaded with factors bind to the 5’ end of the mRNA near the cap. The
LOWRY－PROTEIN－ASSAY
The Lowry procedure is one of the most venerable and widely-used protein assays, being first described in 1951 [Lowry et al., J. Biol. Chem. 193: 265-
Protein－Kinase－A－at－the－Centrosome
Protein kinase A regulatory subunit RIIalpha (PKA-RIIa) is tightly bound to centrosomal structures during interphase through interaction with the A-ki
Bradford－protein－assay
Bradford protein assayConsiderations for useThe Bradford assay is very fast and uses about the same amount of protein as the Lowry assay. It is fairly
BIURET－PROTEIN－ASSAY
BIURET PROTEIN ASSAYMATERIALSBiuret ReagentBovine serum albumin (BSA)Spectrophotometer and tubesPROCEDUREPrepare standard dilutions of BSA containing
Bradford－–－Protein－Determination
Bradford – Protein DeterminationIntroductionA rapid and accurate method for the estimation of protein concentration. The technique is simpler, faster
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