蛋白质技术

Towbin Buffer (5 l)25 mM Tris-Base15.1g192 mM glycine72.1 g20% methanol1 lmake to 5 l with ddH2Odo not adjust the pHTowbin buffer is the same as SDS running buffer except it contains methanol and ...

rehydrate western in PBS-Tween for 5 minadd primary antibody in a minimal amount of PBS-Tween and incubate on a rocking platform for 1 hr min (alternatively overnight at 4C with the tray wrapped in Sa ...

Italics indicate optional steps especially useful for the analysis of untested proteins.GROWTH AND HARVESTING OF BACTERIAAdd 100 ul ampicillin to 100 ml LB. Inoculate and grow overnight.In the morning ...

A. Method for ~1g or more of tissue.1. Label all tubes. Prepare solutions and have ready at hand.2. Remove the tissue from the �80oC freezer and thaw on ice. If the tissue is fresh keep on ice (or alt ...

提起膜蛋白，浮现在广大从事蛋白研究的科研工作者脑海中的是两点：1.膜蛋白功能的重要性。2.膜蛋白研究的困难性。众所周知，膜蛋白在细胞间接触、表面识别、信号转导、酶活性和运输方面都扮演着重要的角色。由于它们功能多样，也就成为理想的药物靶点。然而，膜蛋白的生化和结构研究一直都很缓慢。Protein Database的统计数据表明，在成功解析出三维结构的蛋白中，膜蛋白只占1%，这与膜蛋白占总蛋白的1/3 ...

PurposeThe purification of ICAM-1 on R6.5 anti-ICAM-1 column.Materials Triethylamine NaCl Octyl Glucoside ( n-octyl-b-D-glucopyranoside Sigma Cat. No. O-8001) Tris-Base NaN3 Tris-HCl PMSF (PhenylMethy ...

Purpose MaterialsNUNC-Immuno Plate IIF. Horseradish Peroxidase-Streptavidin (Zymed; 43-4323) PBS or Balanced Salt Solution (BSS). Do not use RPMI or any medium that contains biotin. BSA (1% in PBS). A ...

IntroductionThis is an adaptation of the published procedure for RNAse treatment of Drosophila polytene chromosomes to remove proteins whose binding is RNA dependent such as the dosage compensation co ...

IntroductionHere we describe a strategy for isolation of multiprotein complexes from human HeLa S3 cells in a scale and purity optimized for characterization by mass spectrometry. For this purpose we ...

PROTOCOLTo 1.5 mL eppendorf tubes add:200 µg of protein extract (see Western blot protocol for protein sample preps)q.s. to 300 µL with RIPA (with protease and phosphatase inhibitors).Primary antibody ...

General Preparation1. Prepare buffers and have them cold. Make 50X stock of Buffer G and dilute as needed from frozen aliquots of 50X Buffer G.1X Extraction Buffer without detergents: 1 M Tris-Cl pH 7 ...

co-IP assays can be performed between endogenous proteins or transiently or stably expressed exogenous - usually tagged - proteins. The advantage to using endogenous proteins is the avoidance of prote ...

Western Blot Workflow Diagramback to topProtocolStandard vs. Rapid Immunodetection ProceduresThere are two types of protocols for immunodetection: Standard and rapid. Standard vs. Rapid Immunodetectio ...

1) After ECL development wash membrane once for 10min with PBST.2) Incubate the membrane in stripping buffer (see below) in a heat-sealed plastic bag for 30min at 50oC with occasional mixing. For low- ...

Introduction After proteins have been separated by electrophoresis individual protein bands can often be identified by using an antibody that is specific to that protein. However to be accessible ...

INTRODUCTION In 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 incub ...

INTRODUCTION In 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 factor Xa temperature and length of incu ...

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) Take competent cell stock aliquot (about 100 mL) o ...

MaterialsRIPA buffer (RIPA buffer enables the extraction of cytoplasmic membrane and nuclear proteins and is compatible with many applications including reporter assays protein assays immunoassays and ...

OverviewThis is a protocol for preparing whole cell lysate from tissue for western blot analysis. MaterialsRIPA Bufferper 100ml: (final concentration) 5ml of 1M Hepes pH 7.6 (50 mM) 200µL of 0.5M EDTA ...