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We use two different kinds of media. Most cells are grown in DMEM. A few lymphoid cell lines are grown in RPMI. Cells grown in DMEM must be grown in a 10% CO2 atmosphere. As a result, most of our incubators are set for this. In contrast, cells grown in RPMI must be grown in a 5% CO2 atmosphere. We have one incubator set up for this. If you grow cells in RPMI in 10% CO2, the medium will be too acidic (yellow).DMEM.Thi......閱讀全文
MaterialsFibroblast suspension cultureTissue culture laminar flow hoodMedia appropriate to culture line usedDisposable pipettes (10 ml and 1.0 ml)Disp
· Cell Culture Media and Solutions (Donis-Keller lab)· &n
REFERENCES:R. Ian Freshney, Culture of Animal cells: A manual of basic techniques, Wiley-Liss, 1987.VI. TISSUE CULTURE PROCEDURESEach s
AimDNA staining methods such as Hoechst staining techniques are quick with results available within 24 hours, which compares favorably with 4 weeks fo
General StrategyWe typically work with tissue culture, primary mammalian cells, and cell extracts, but the protocols can be adapted to other systems,
Fluorescence Procedures for the Actin and Tubulin Cytoskeleton in Fixed CellsActin: Louise CramerTubulin: Arshad DesaiGeneral StrategyWe typically wor
Exercise 12.10 - Establishment of a Primary CultureLEVEL IIIMaterialsChick embryo (approximately 8 days old)70% (v/v) ethanol for swabbingSterile scis
實驗概要This Rat/Mouse Growth Hormone ELISA kit is used for the non-radioactive quantification of Growth Hormone in rat or mouse serum, plasma
I. Purpose:A. Skin tissue may be used for chromosome analysis in special cases when the results from peripheral blood are inconclusive, e.g. susp
實驗概要 Skin tissue may be used for chromosome analysis in special cases when the results from peripheral blood are inconc
Method: Cell Counts Using a HemacytometerJune 1, 1990Rosalie VeilePurpose:The purpose of this procedure is to determine the cell density of the cultur
I. Purpose:A: Skin tissue may be used for chromosome analysis in special cases when the results from peripheral blood are inconclusive, e.g. suspected
Primary Culture of Pulmonary Arterial Smooth Muscle Cells 1.Primary cultures of Pulmonary Arterial Smooth Muscle Cells (PASMCs) were isolated fro
Picking ES cell clonesOne or two days before picking colonies prepare 24-well plates of feeders. You can also use alternate protocols that utilize 96-
常規操作(主要內容如下)· Aseptic Technique· Culture Ves
When an irreplaceable culture becomes contaminated, researchers may attempt to eliminate or control the contamination. First, determine if the contami
Early in embryonic development, the region of the chick embryo which is determined to form a limb first differentiates from the rest of the embryo1. T
OverviewMatirgel is considered as basement membrane and generated from EHS sarcoma. Matrigel contains not only basement membrane components (collagens
Purpose:This method is advantageous for saving the occasional cultures that become contaminated. Yeast contaminated cultures will appear cloudy when s
（3）NE+IFN-DMEM（-6M，50ng）NE---A液（1μg/μl，1mg/ml） 20.5μlIFN-γ（25ng/μl） 25 μlDMEM UP TO 100 ml過濾消毒，4℃保存（4）低血清DMEM培養基（上室）（5）20%FBS-DMEM培養基（下室）2、準備（1）溶膠，4℃過
MaterialsTrypsin (Gibco 25200-023)3T3 Medium: 500 mL DME (Invitrogen) + 50 mL FBS (Hyclone) + 5 mL 100x Pen/Strep2x Freezing Medium: 3T3 Medium
一 、 原理Matrigel 是從小鼠EHS肉瘤中提取的基質成分，含有LN、IV型膠原、接觸蛋白和肝素硫酸多糖，鋪在無聚乙烯吡硌烷酮的聚碳酸酯濾膜上，能在DMEM培養基中重建形成膜結構，這種膜結構與天然基質膜結構極為相似。濾膜孔徑一般為8um，而且膜孔都被Matrigel覆蓋，細胞不能自由穿過，必須
We use feeder-independent ES cell lines derived from the 129/Ola strain of mice (Nichols et al., Development 110, p.1341, 1990). These cells are easy
Noninvasive Human Nuclear Transfer with Embryonic Stem CellsSohyun L. McElroy1 and Renee A. Reijo PeraCenter for Human Embryonic Stem Cell R
PurposeThe purpose of a chemotaxis assay is to determine whether your protein or small molecule of interest has chemotactic activity on a specific cel
Springer Lab，The CBR Institute for Biomedical Research, Inc. Department of Pathology Harvard Medical Schoolhttp://cbr.med.harvard.edu/investigato
MaterialsRIPA buffer (RIPA buffer enables the extraction of cytoplasmic, membrane and nuclear proteins and is compatible with many applications, inclu
Isolation of renal papillary cells1. For isolation of papillary cells, kidneys were harvested and kept in HBSS containing 15 mM HEPES, penici
Fine Mapping of Genomic Targets of Nuclear Proteins in Cultured CellsAchim Breiling and Valerio OrlandoDulbecco Telethon Institute, Institute of Genet
In vitro culture of embryonic lungsfrom Hogan LabIsolation of Lung Bud EndodermWhat you need:E11-12 mouse embryosDMEM with 5% fetal bovine serumpetri