Growth,MaintenanceandTransfectionofSuspensionAdapted293EBNAcells

ProcedureI. INTRODUCTIONThe 293 EBNA cell line is established from primary embryonal human kidney cells transformed with sheared human adenovirus type 5 DNA and Epstein-Barr virus nuclear antigen 1, allowing episomal amplification of plasmids containing the viral EBV origin of replication. This protocol describes the instructions for the growth, maintenance and transfection of suspension adapted 293-EBNA cell line us......阅读全文

Growth,-Maintenance-and-Transfection-of-Suspension-Adapted-293EBNA-cells

ProcedureI. INTRODUCTIONThe 293 EBNA cell line is established from primary embryonal human kidney cells transformed with sheared human adenovirus type

Transfecting-Suspension-Cells

实验概要将转移基因整合到细胞染色体DNA上,形成稳定表达转移基因的细胞系。 实验原理    细胞转染技术是目前广泛应用于病毒基因结构与功能以及基因调控等的研究。细胞转染可分为短暂转染和稳定(或永久) 转染两种。在短暂转染中,被转染基因并不整合至细胞染色体中,因而不能随细胞分裂而传代。转入病毒基因的转

Selection-of-Transfected-Suspension-Cells

Contributor: Suprya JayadevDate: December 13, 19941) Transfect cells.2) Culture cells 1-3 passages in a T-75 flask containing selection material (e.g.

Transfection-of-Mammalian-Cells-Using-Lipofectamine

Materials:       LipofectamineBasal Medium containing 10% fetal bovine serum, 1% glutamine, 1% aaBasal Medium containing 1% glutamineBasal Medium cont

Transient-Transfection-of-Cos1-Cells

Transient Transfection of Cos-1 CellsNote: This protocol has been optimized for Cos-1 cells. Successful transfection of each cell type requires optimi

Transient-transfection-into-293T-cells

PurposeTransient transfection into 293T cells is a convenient way to overexpress and obtain both cellular and extracellular (secreted or membrane) pro

293fectin™-Transfection

实验概要293fectin™  is a proprietary, cationic lipid-based formulation for transfection of  DNA into eukaryotic cells. 293fectin™ is optimized for transfe

96Well-Sample-Preparation-for-Suspension-Cells

实验概要The procedure  presented below describes a facile method for studying signal  transduction events with suspension cells (Jurkat, Raji, THP-1, etc.

96Well-Sample-Preparation-for-Suspension-Cells

实验概要The procedure  presented below describes a facile method for studying signal  transduction events with suspension cells (Jurkat, Raji, THP-1, etc.

96Well-Sample-Preparation-for-Suspension-Cells

实验概要The  procedure presented below describes a facile method for studying signal  transduction events with suspension cells (Jurkat, Raji, THP-1, etc.

DNA转染

DNA转染·         Transfection of Mammalian Cells Using Lipofectamine (LTI)·         Guide to Eukaryotic Transfections with Cationic Lipid Reagents (PDF)

Fusion-and-Cloning

ReagentsMedium A - Pre-fusion Medium and Hybridoma Expansion MediumMedium B - Fusion Medium Medium C - Hybridoma Recovery MediumMedium D - Hybridoma S

PriCells:-Questions

 Questions 1: Isolation of hepatocytes from other speciesThe basic two-step perfusion procedure can be used for obtaining hepatocytes from various r

CO2恒温摇床解决人胚肾-293-(HEK293)-细胞结团问题(二)

Lysate preparation and western blottingProtein lysates were created by harvesting the cells from confluent T-flasks or from suspension cultures at h

Growing-Cells-in-Geltrex™-Reduced-Growth-Factor-Basement-Membrane-Matrix

实验概要Basement  membranes are continuous sheets of specialized extracellular matrix  that form an interface between endothelial, epithelial, muscle, or

哺乳动物RNAi技术-Mammalian-RNA-Interference

Mammalian RNA InterferenceThomas TuschlLaboratory for RNA Molecular BiologyThe Rockefeller University, New York Excerpted from RNAi: A Guide To Gene S

细胞培养常规操作

常规操作(主要内容如下)·         Aseptic Technique·         Culture Vessels·         Cell Counting·         Primary Culture·         Maintenance of Cell Line ·  

Isolation-of-human-endometrial-epithelial-cells

Tissue collection1. Endometrial biopsies were collected from women undergoing gynaecological procedures for benign conditions. 2. All women reported

Fibroblast-Cell-Systems3

Seeding After cells are thawed:NOTE: Do not dispense the entire contents of the cryovial into one T-25 flask!!Remove the cap, being careful not to tou

使用CO2恒温摇床解决人胚肾-293-(HEK293)-细胞结团问题

人胚肾 293 (HEK293)  细胞在重组蛋白表达中是最常见的宿主细胞。 这类细胞能够表达大量的膜蛋白,如 G 蛋白偶联受体  (GPCR) ,是无法在最常见的生物制药生产宿主,如:中国仓鼠卵巢 (CHO) 细胞中作表达。 HEK293 虽然是蛋白表达的极好宿主,然而 HEK293 细胞

使用CCCadvanced™FN1无异源耗材培养人多能干细胞(二)

Materials and MethodsCell culture conditions and surface transitionCryopreserved hiPSCs (SC102A-1, SBI™, USA) were initially thawed and pre-cultivat

Maintenance-of-Cell-Culture

Maintenance of Cell CultureAuthor: Nanci DonackiSource: Contributed by Nanci DonackiDate Added: Tue May 14 2002Date Modified: Tue Apr 27 2004Abstract:

Fusion-and-Cloning

Author: Nanci DonackiSource: Contributed by Nanci DonackiAbstract: Procedure for establishing hybridoma in one stepReagents(StemCell Technologies, Inc

Tissue-Culture-Methods3

REFERENCES:R. Ian Freshney, Culture of Animal cells: A manual of basic techniques, Wiley-Liss, 1987.VI. TISSUE CULTURE PROCEDURESEach student should m

Transfecting-Plasmid-DNA-into-NIH3T3-Cells-Using-Lipofectamine™-LTX-Reagent

实验概要Lipofectamine™  LTX Reagent is a proprietary, animal-origin free formulation for the  transfection of DNA into eukaryotic cells with low cytotoxic

Fibroblast-Cell-Systems4

3. If your result falls into any quadrant other than the "High Yield-High Viability" quadrant, refer to Appendix D, Improving Cell Yield and Viability

Rat-urinary-bladder-urothelial-cells

1. Bladders were excised from deeply anesthetized (urethane, 1.2 gm • kg−1, i.p.) Sprague Dawley rats (of either sex), cut open, and gently stretc

Handling-Fresh-Hepatocytes-in-Suspension

实验概要Important—Please read!GIBCO®  Fresh Hepatocytes are shipped in cold preservation medium designed to  keep cells viable at 4°C. This shipping mediu

Transfection-with-PEI

实验概要Use PEI (Linear 25 kDa Reagent) to transfect in HEK293T cells.主要试剂PEI is polyethyleimine, a 25 kDa linear from Polysciences Inc. Make up solutio

Fibroblast-Cell-Systems6

APPENDIX E GROWTH AREA OF COMMON PLASTICWAREFlasksEffectiveGrowthAreaInitial Number of Cells to Seed at 3500 cells/cm2 NHDFExpected Number of Cells at