SEMIDRYELECTROPHORETlCTRANSFER(WESTERNBLOTS)
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 to antibodies the proteins in the gel must first be transferred from the gel to a membrane. This process is called "Western blotting" (after E. M. Southern). We will use the &q......阅读全文
SEMIDRY－ELECTROPHORETlC－TRANSFER－(WESTERN－BLOTS)
Introduction After proteins have been separated by electrophoresis, individual protein bands can often be identified by using an antibody that is
Southern－Blotting:－DNA－Transfer
Southern Blotting: DNA Transfer1. Depurination of DNA fragments: Wash gel in 0.25 M HCl 5' (small gels), 7' (large gels)2. Denaturation: Wash
Transfer－of－Eukaryote－Suspension－Cultures
MaterialsFibroblast suspension cultureTissue culture laminar flow hoodMedia appropriate to culture line usedDisposable pipettes (10 ml and 1.0 ml)Disp
Horizontal－Transfer－of－Supernumerary－Chromosomes－in－Fungi
Several species of filamentous fungi contain so-called dispensable or supernumerary chromosomes. These chromosomes are dispensable for the fungus
Dry－Transfer－干法蛋白转膜
Trans-Bot SD Assembly1. Prepare the transfer buffer.2. Following electrophoresis, equilibrate the gels in transfer buffer. Equilibration facilitates
Shuttle－for－transfer－of－acetyl－groups－from－mitochondria－to－the－cytosol
Acetyl-CoA is produced in mitochondria through the metabolism of fatty acids and the oxidation of pyruvate to acetyl-CoA. When ATP is needed, this ace
Noninvasive－Human－Nuclear－Transfer－with－Embryonic－Stem－Cells
Noninvasive Human Nuclear Transfer with Embryonic Stem CellsSohyun L. McElroy1 and Renee A. Reijo PeraCenter for Human Embryonic Stem Cell Research an
BioRad（伯乐）Western－Blot半干法转膜的十大注意事项
首先讲转膜仪的清洗：Do not immerse the unit in liquid. Use special care when cleaning the anode plate to avoid scratching or marring the platinum. Do not use
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
Western－杂交
Western 杂交（主要内容如下）Preparing of Protein LysatesWestern BlottingFar Western BlottingSemi Dry BlottingStripping MembranesTrouble Shooting and OthersPrepa
Immunoblotting－(Western－Blotting)
实验概要We provide a protocol for SDS-PAGE, Protein Blotting, Immuno-Detection.主要试剂1. 0.3 M TRIZMA® base (Product No. T1503), 20% methanol.2. 0.025 M TRIZ
Western－Blot－(免疫印记法)常识问答（二）
4. western blot 所需buffer 的配方是什么？答：主要的buffer有：1） 2×Separation buffer (PH: 8.8)0.75M TRIS.HCl 90.825g0.2% SDS 10ml 20% SDSTotal: 1000ml2) 30% gel Soluti
免疫印记(WB)常见问题解答
1. 什么是western blot?答：WB又称免疫印记法.是将生物大分子物质通过不同途径转移到固相载体的过程。 2. western blot 有什么优点？答：灵敏，可达ng级，用Ecl显色法理论上可达pg 级。方便，特异性高。 3. western blot 的具体步骤是什么？答：一. 制样（
Western－Blot－(免疫印记法)常识问答
1. 什么是western blot? 答：WB又称免疫印记法.是将生物大分子物质通过不同途径转移到固相载体的过程。 2. western blot 有什么优点？ 答：灵敏，可达ng级，用Ecl显色法理论上可达pg 级。方便，特异性高。 3. western blot 的
Western－Blot－(免疫印记法)常识问答
1. 什么是western blot? 答：WB又称免疫印记法.是将生物大分子物质通过不同途径转移到固相载体的过程。 2. western blot 有什么优点？ 答：灵敏，可达ng级，用Ecl显色法理论上可达pg 级。方便，特异性高。 3. western blot 的
Typical－Western－Tra...
实验概要Peprotech provides a typical western transfer and development protocol.实验原理Western Transfer, also known as Western Blotting, is a rapid immunobl
COLONY－HYBRIDIZATION
COLONY HYBRIDIZATION1) CUT 4 PIECES OF 3MM WHATMAN PAPER AND PLACE EACH ONE IN A SEPARATE CONTAINER(A CAFETERIA TRAY WILL PROBABLY WORK PERFECTLY).2)
SOUTHERN－BLOT的步骤
1. Run the gel as normal. Often for genomic southerns it is desirable to run long gels (18cm) over 4-6hrs.2. Photograph the gel with a ruler adjacent
Southern杂交技术
SOUTHERN BLOT1. Run the gel as normal. Often for genomic southerns it is desirable to run long gels (18cm) over 4-6hrs.2. Photograph the gel with a r
Coimmunoprecipitation－assays
co-IP assays can be performed between endogenous proteins or transiently or stably expressed exogenous - usually tagged - proteins. The advantage to u
Immunoblot－Protocol
This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor
In－vitroculture－of－early－chick－embryos
1. Use sterile technique. Prewarm Howard's Ringers in petri dish andagar/albumin culture dish to 37ºC.2. Crack 2-day egg into large sterile petri
Southen杂交
Southern杂交One important thing for transfer:the weight of the object resting on top of the blotting apparatus should not exceed the weight equal to a 5
Smolke:Protocols/Western
OverviewBlotting for large V5-tagged proteins in S. cerevisiaeMaterialsY-PER (Pierce)Halt EDTA-free Protease Inhibitor (Pierce)NuPAGE Novex Bis-Tris 4
Cryopreservation－of－cell－cultures
1. Examine all flasks by inverted-phasemicroscopy. Cultures used for preservation should be grown free of antibiotics, show no signs of microbial cont
Preparation－of－Genomic－DNA－from－Bacteria－using－Phase－Lock－GelTM
Preparation of Genomic DNA from Bacteria- using Phase Lock GelTM (Modified from Experimental Techniques in Bacterial Genetics, Jones and Bartlet, 1990
DNA－Extraction－from－Tissue
实验概要DNA extraction from tissue.主要试剂Extraction buffer100 mM Tris-HCl (pH 8.0) 100 mM EDTA (pH 8.0) 100 mM Na-Phosphate (pH 8.0) 1.5 M NaCl1% CTAB
UV－CrossLinking－an...－（二）
实验步骤1. UV cross-linking of tissue culture cells 1) Remove media and add ice-cold PBS to cells (e.g. use cells grown in a 10 cm plate for three ex
Removing－cells－from－liquid－nitrogen
Put cryovial straight from storage and float in the 37篊 water bath- caution should be taken as on rare occasions vials can explode when heated up due
分子生态学词汇水平基因转移
中文名称：水平基因转移外文名称：Horizontal gene transfer又 称：基因侧向转移类 型：遗传学术语定义：基因水平转移（英语：Horizontal gene transfer，HGT），又称水平基因转移或基因侧向转移（英语：lateral gene transfer，L