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
Fusion－and－Cloning
Author: Nanci DonackiSource: Contributed by Nanci DonackiAbstract: Procedure for establishing hybridoma in one stepReagents(StemCell Technologies, Inc
Fusion－and－Cloning
ReagentsMedium A - Pre-fusion Medium and Hybridoma Expansion MediumMedium B - Fusion Medium Medium C - Hybridoma Recovery MediumMedium D - Hybridoma S
Quick－Yeast－DNA－Prep:－Isolation－of－Total－DNA－(genomic－and－plasmid)
Grow a 5 ml YPD O/N culture inoculated with a single yeast colony at 30 deg.Transfer culture to a small 13 x 100 glass tube. Spin down cells 2 min. in
Partial－Endonuclease－Digestion
Partial Endonuclease DigestionPrepare a 100 µl reaction mixture containing the DNA of interest in the appropriate 1X restriction enzyme buffer. Divide
Phage－DNA
IntroductionThe phage lysate from the plate contains bacterial DNA and RNA, as well as phage DNA encased in the phage coat. The following procedure, d
Replica－Printing
Jeff Lawrence (M05)1. Sterilize Velvet Squares with widths at least 50% longer than the diameter of a petri plate.2. Streak strains to be printed for
Method:－Preparation－of－Lymphocyte－Cell－Pellet－for－Storage
Method: Preparation of Lymphocyte Cell Pellet for StorageJune 10, 1990Rosalie VeilePurpose:Following propagation to 1 X 108 cells, lymphoblastoid cell
DNA－and－RNA－EXTRACTIONS
A protocol / method / schedule /procedure for extraction / isolation of both DNA and RNA from the same material typically plant leaf / leaves(See also
Cryopreserving－Neural－Stem－Cells
实验概要There are numerous protocols available for cryopreserving neural stem cells (NSCs) derived from human embryonic stem cells; the primary objectiv
DNA－EXTRACTION－PROCEDURE－－GENERAL
Grow cells overnight in 500 ml broth medium.Pellet cells by centrifugation, and resuspend in 5 ml 50 mM Tris (pH 8.0), 50 mM EDTA.Freeze cell suspensi
Maintenance－of－Cell－Culture
Maintenance of Cell CultureAuthor: Nanci DonackiSource: Contributed by Nanci DonackiDate Added: Tue May 14 2002Date Modified: Tue Apr 27 2004Abstract:
Sphingomyelin－Quantitation－Postcholine－Labeling－of－HL60－Cells
Lipid Extraction1) Following the appropriate time of treatment, transfer 4.5 ml into each of two duplicate glass pyrex tubes and maintain on ice.2) Sp
Culturing－Mouse－Embryonic－Fibroblasts
MaterialsTrypsin (Gibco 25200-023)3T3 Medium: 500 mL DME (Invitrogen) + 50 mL FBS (Hyclone) + 5 mL 100x Pen/Strep2x Freezing Medium: 3T3 Medium + 20%
Tube－formation－assay
DescriptionThis is a fast and easy assay to test the angiogenic/anti-angiogenic properties of molecules. As compared to other angiogenesis assays, suc
分子生态学词汇遗传传递
中文名：遗传传递外文名：genetic transfer传递方式：转化、转导和接合特 征：传递过去的基因部分部分接合子定义：遗传传递 genetic transfer 专指细菌中遗传物质在细胞间的传递。其传递方式包括转化、转导和接合等。但仅在接合的情况下，有极少数的供体菌的全部基因组可传递给受
Nucleofection
This is an extract of the Amaxa Biosystems protocol Vs. 09-2005 optimized for use with the UC06 cell line. It is suggested that you try all 5 programs
Method:－Lymphoblastoid－Cell－Lines－from－Frozen－Whole－Blood
Method: Lymphoblastoid Cell Lines from Frozen Whole BloodMay 31, 1990Rosalie VeilePurpose:Blood Samples can be stored frozen as a backup in case an LC
Sea－Urchin－Fertilization
Gametes are collected as described. To remove some of their jelly coat,wash eggs several times in ASW by allowing them to settle and gently pouring of
2D－Polyacrylamide－Gel－Electrophoresis
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
潘远江教授：电喷雾质谱中的苄基迁移反应研究
浙江大学 潘远江教授 2014年4月26日，首届全国质谱分析学术研讨会在北京西郊宾馆盛大开幕。来自浙江大学的潘远江教授带来了题为《电喷雾质谱中的苄基迁移反应研究》的报告。 潘远江教授首先介绍了他们课题组的气相离子化学研究：解离质子化位点理论模型；苄基阳离子迁移理
酵母染色体沉淀分析方法
ABSTRACTThis protocol describes a method for the detection of proteins bound to specific regions of chromatin in yeast. There are many variations of t
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
Pouring－Plates
1. For rich media, weigh out appropriate ingredients and place into a flask. Add water until appropriate volume. Use a flask at least 2 times larger t
细胞膜蛋白质提取方法
NRC Institute for Biological SciencesTriton X-114 extraction protocol (Hydrophobic protein preparation)Ressuspend cells in Solution A (dil 1/8) and ad
Cryopreservation－and－Recovery－of－Mature－Differentiated－Neural－Cells
实验概要Primary neuronal cultures are indispensable in the field of neurobiology and pharmacology. Many researchers favor freshly isolated neuronal cell
Western－blotting－Protocol(试剂配方和实验操作)
一、 试剂配方：1X TBS: Tris-base 12.11gNacl 8.775g用HCl 调pH7.4，用纯水稀释至1L5X Transfer buffer: Tris-base 15.1gGlycine 72.0gTo 1 L10X碱性磷酸酶缓冲液： 1 mol/L Tris-HCl pH
Thawing－Cells－(Schreibers－protocol)
Thaw vial quickly in 37癈 water. Caution - vial can explode.Transfer cells to sterile, 15 mL centrifuge tube.Add 50 祃 warm FBS (fetal bovine serum, hea
Preparation－of－fixed－embryos－for－immunocytochemistry－and－AP－staining
1. Transfer 50 ml of embryo cultures to centrifuge tubes. Spin at 1500 rpm for 5 minutes. Check that you can see a pellet of embryos at the bottom.Qui
Isolation－of－Zebrafish－embryos
Zebrafish will mate and deposit fertilized eggs on the bottom of the tank at 'dawn'. They can be accustomed to lay at any convenient time by k
SOLID－TUMOR－CULTURES－FOR－CHROMOSOME－ANALYSIS
I. Purpose:A. Samples of solid tumors or lymph nodes may be sent from patients with cancer. These samples should be processed directly and also set up