ProtocolforCellFusion
Healthy Sp2/0 cells should be rapidly growing by this time. Sp 2/0 cells should be started about two weeks before the cell fusion. Every two days, they should be centrifuged at a 64.4 xg on the IEC clinical centrifuge (in 50 ml tubes) for two minutes. The media should be removed and the cells resuspended in fresh media. This ensures that only the healthiest cells will fall to the bottom of the tube. Centrifugation at......阅读全文
DNA－Cell－Cycle
Solutions70% ethanolribonuclease (100 µg/ml DNase free, Sigma)propidium iodide ( 50 µg/ml in PBS)ProcedureHarvest cells. Spin at 1200 rpm for 5 minute
Apoptosis－Induction
IntroductionWhen studying induction of apoptosis via a cell surface molecule, it is important to first ascertain surface expression of the molecule of
Extraction－of－DNA－using－DNAzol®－Reagent
实验概要DNAzol® Reagent (Genomic DNA Isolation Reagent) is a complete and ready-to-use reagent for the isolation of genomic DNA from solid and liquid sa
Human－Embryonic－Stem－(ES)－Cell－Protocols——Splitting－Human－ES－cells－on－MEFs
based on splitting onto one plateWarm collagenase IV split media to 37 °C in a water bath.Aspirate media off of cell culture plate.Add the following a
Human－Embryonic－Stem－(ES)－Cell－Protocols—Splitting－Human－ES－cells－onMatrige
based on splitting onto on plateWarm collagenase media to 37°C in a water bath.Aspirate media off of cell culture plate.Add the following amount of co
Inoue法制备大肠杆菌超级感受态细胞
实验步骤： 1、Inoculate from an overnight grown in LB.从培养过夜的LB平板上挑取单菌落 。2、Grow in 250 ml "SOB" at 18℃ until OD600 = 0.6.(0.3)接种于250ml SOB，18度培养至OD＝0.6。3、On
PBMC细胞的精确计数和活性分析（三）
五、使用仪器发表文章AuthorDateTitleJournalCell TypeCellometer / ApplicationsMahato, Ram INovember 2013Synthesis and Characterization of an Anti-Apoptotic Immu
Soft－Agar－Assay
Soft Agar AssayMake 0.6% media-agar mix for the bottom layer. To make 0.6% agar mix the following components (this makes 200 ml):2X DME 100 mlI
Trypsinizing－cells
There are many procedures with which to trypsinize cells. All include washing the cell monolayer with TD, or in rare cases, with VE. This removes seru
Combined－Flow－Cytometric－Measurement－of－Two－CellSurface－Antigens
INTRODUCTIONFlow cytometry is frequently used to assess nucleic acid content in individual cells. Based on DNA content alone, however, cells in the qu
Human－DC－Enrichment－Kit
实验概要Enrich untouched Dendritic Cells (DCs) by depleting T cells, B cells, monocytes/ macrophages, NK cells, erythrocytes and most granulocytes from
Preparing－Lambda－DNA
Preparing Lambda DNA1- Coliphage lambda DNA is a widely used vector for recombinant DNA. The middle third of its 48,000 bp contains no genes required
Infection－with－retroviruses
It is well established with avian retroviruses that cells are most efficiently infected just after they are trypsinized. Trypsinization does two thing
亲和层析实验技术方法
INTRODUCTIONThis protocol describes a method for removing antibodies that react with bacterially encoded proteins by passing a crude preparation of im
体外荧光法检测核内体早期动力学5
Leave tubes on ice and repeat Steps 13–15 for the next 6–12 plates of cells.When all cells are collected, centrifuge all tubes at 250g for 5 min at 4
Measurement－of－Green－Fluorescent－Protein－Expression
ReagentsCells to be studied expressing green fluorescent protein (GFP). Note that the same cell type without GFP is needed as control.Hoechst 33342 s
Isolation－of－retinal－pigment－epithelial－cell（二）
15. This medium is replaced after 1 day with 5% serum-containing RPE medium, and subsequent changes were made every 2 to 3 days.16. After 3 to 4 weeks
High－Efficiency－Transformation
Day 0Make sure you have the necessary solutions (instructions for how to make them can be found here):Single-stranded carrier DNAPEG 3350 50% w/vol1.0
Critical－Appraisal－of－the－MTT－Assay－in－the－Presence－of－Rottlerin－6
Our experience indicates that it may not be sufficient to change the medium containing Rottlerin and to wash the cells before adding MTT to avoid a po
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
Staining－Methods－for－cell－death－Z.－Xia－10/2/95
The simplest way: trypan blue. Dead cells stain blueNon-fixed cells: FDA(fluorescein diacetate)-green, alive cells; P.I. (propidium iodide)-red, dead
Staining－Methods－for－cell－death
The simplest way: trypan blue. Dead cells stain blueNon-fixed cells: FDA(fluorescein diacetate)-green, alive cells; P.I. (propidium iodide)-red, dead
Staining－Methods－for－cell
death Z. Xia 10/2/95The simplest way: trypan blue. Dead cells stain blueNon-fixed cells: FDA(fluorescein diacetate)-green, alive cells; P.I. (propidiu
Agrobacterium－growth－and－transformation
Growth and storage of Agrobacterium tumefaciensStrain GV3101: resistant to gentamycin and rifampicin so add 25-50 ug/ml Gentamycin, 10 ug/ ml rifampic
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
DAPI－Counterstaining－Protocols
实验概要The blue-fluorescent DAPI nucleic acid stain preferentially stains dsDNA; it appears to associate with AT clusters in the minor groove. Binding
A－Method－for－Assaying－Deubiquitinating－Enzymes1
AbstractA general method for the assay of deubiquitinating enzymes was described in detail using 125I-labeled ubiquitin-fused αNH-MHISPPEPESEEEEEHYC (
Realtime－PCR
实验概要The exponential amplification via reverse transcription polymerase chain reaction provides for a highly sensitive technique in which a very low
Fluorescence－Procedures－forthe－ActinandTubulin－Cytoskeleton－inFixed－Cells1
General StrategyWe typically work with tissue culture, primary mammalian cells, and cell extracts, but the protocols can be adapted to other systems,
Xenograft－Tumor－Model－Protocol
Preparation of tumor cellsGrow cells in complete medium and exclude any contaminationWhen cells are 70-80% confluent, 3-4 hrs before harvesting, repla