[3H]ThymidineIncorporationAssayforRat1acells
Overview This method of Peter Coward, Ph.D. in the Conklin Lab was used in Coward, et al (1998) Controlling signaling with a specifically designed Gi-coupled receptor Proc. Natl. Acad. Sci. 95:352-357.(http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?uid=9419379&form=6&db=m&Dopt=b)More information from the Conklin Lab is available at (http://gladstone.ucsf.edu/conklin.html) Mater......阅读全文
GST－Activity－Fluorometric－Assay
实验概要The experiment provides a simple, fluorescence-based in vitro assay for detecting the GST activity using a fluorescence plate reader. The assay
Cell－Clonogenic－Survival－Assay
DescriptionAllows one to test the capability of adherent cells to survive and replicate following insult with chemicals or radiation. Procedure1. Grow
Hanging－drop－aggregation－assay
DescriptionThis assay is used for the aggregation property of cancer cells. It is a very critical parameter for measurement of cell metastasis. Factor
Alanine－Transaminase－Activity－Assay
实验概要Alanine Transaminase (ALT) is a transaminase (EC 2.6.1.2) also called serum glutamic pyruvic transaminase (SGPT). Alanine Transaminase is found
Phosphate－Assay－by－Suprya－Jaydev
ReagentsAshing buffer:10 g Mg(NO3)2 100 ml EtOH1.5 N HCl stock:119.7 ml concentrated HCl (11.6M @ 36% by weight)880.3 ml H2O1 N Sulfuric acid stock:28
AlamarBlue®－Cell－Viability－Assay
实验概要Assess cell viability. 实验原理Cell health can be monitored by numerous methods. Plasma membrane integrity, DNA synthesis, DNA content, enzyme activ
Enzyme－Kinetics－assay－of－the－WT
To assay 17 b-HSD activity in lysates, cells were harvested 48h after transfection using PBS Enzyme Free Cell Dissociation Solution ( specialty Media
MTT－Cell－Proliferation－Assay
MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay, first described by Mosmann in 1983, is based on the ability of a mitochondri
Gel－Shift－Assay－Systems
ProtocolsDownloadprotocol183kbpdf?Abstract for Gel Shift Assay SystemsThe gel shift, or electrophoretic mobility shift, assay provides a simple and ra
Polyphenoloxidase－(catechol－oxidases)－assay
Browning of the cut surface of some fruits and vegetables is due the presence of a group of enzymes called polyphenoloxidases. These enzymes are relea
In－Vitro－Protein－Ubiquitination－Assay
Ubiquitination is one of the most important posttranslational modifications in all eukaryote organisms. Ubiquitin-activating enzyme (E1), ubiquiti
Chorioallantoic－Membrane－(CAM)－Assay
8 eggs per day, day 7- day 13 cut CAM, wash in precooled PBS,in 10 ml WASH 1 (PBS, 5 mM EDTA, COMPLETE) on icecut in pieces in petri dish on icecentri
Nucleotide－Binding/Hydrolysis－Assay
MaterialsNucleotide mixMotor (50 - 100 µM; purity > 95%)0.5 M Tris-OAc, pH 7.510 mM EGTA10 mM MgCl2DDWSephadex G-50 Medium column (0.8 cm in x 20 cm)C
Preparing－cells－and...
实验概要The method provides a protocol and tips for BrdU staining in tissue sections.Bromodeoxyuridine (5-bromo-2-deoxyuridine, BrdU) is a synthetic
Collection－of－Peritoneal－Cells
Prepare a 10ml syringe fitted with a 26G short needle and filled with 5 to 7 ml of medium and 2 to 3 ml of air. Air is important.Prepare a Pasteur pip
Collection－of－Peritoneal－Cells
Prepare a 10ml syringe fitted with a 26G short needle and filled with 5 to 7 ml of medium and 2 to 3 ml of air. Air is important.Prepare a Pasteur pip
Fluorescent－Staining－of－Cells
1. Fluorescent phalloidin in methanol. Phallacidin does not work as well. Dilute 10 ul 330 nM stock into 500 ul PBS for each large coverslip.2. PB
Freezing－and－Thawing－cells
Freezing and Thawing cellsFreezingIt is best to freeze cells that are growing rapidly. With adherent cells, it is easiest to set up 100 mm dishes, giv
Subculturing－Adherent－Cells
实验概要The following protocol describes a general procedure for subculturing adherent mammalian cells in culture.主要试剂1. Complete growth medium, pre-warme
Immunofluorescence－Labeling－of－Cells
实验概要Antibodies are an important tool for demonstrating both the presence and the subcellular localization of an antigen. Cell staining is a very ver
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
KARYOTYPING－ES－CELLS
An actively growing culture of cells is required, i e 2 - 3 d ES cell culture. The total number of cells needs to be between 106 - 107 cells.N B Read
Electroporation－of－ES－cells
Cells are routinely passaged two days prior to electroporating. Usually one 10 cm plate at approximately 80% confluency will provide enough cells for
Isolation－of－papillary－cells
实验概要This protocols provides a general protocol for isolation of papillary cells.实验步骤Isolation of renal papillary cells1. For isolation of papillary c
Isolation－of－papillary－cells
Isolation of renal papillary cells1. For isolation of papillary cells, kidneys were harvested and kept in HBSS containing 15 mM HEPES, penicillin/
DNA转染
DNA转染· Transfection of Mammalian Cells Using Lipofectamine (LTI)· Guide to Eukaryotic Transfections with Cationic Lipid Reagents (PDF)
Invitro－Phagocytosis－Assay－of－Macrophages
IntroductionThe term phagocytosis itself describes its mean phage = engulfment; cytosis: cell process. In other words, phagocytosis is the cellular pr
Chick－Chorioallantoic－Membrane－(CAM)－Assay
CAM ASSAYShell-less embryo cultureFertilized white leghorn chicken eggs (SPAFAS Inc., Norwich, CT) were received at day 0 andincubated for 3 days at 3
Apoptosis－TUNEL－Assay－(frozen－sections)
Protocol for Frozen Sections:Warm 150ml 4% Paraformaldehyde/1x PBS to RT. Fix slides in it, 20 min., RT.1x PBS rinse, 2 times.1x PBS, 30 min., RT. Beg
The－UnderAgarose－Migration－Assay
overviewThe Under-Agarose assay is a useful method for observing the response of a cell population to one or more chemoattractant sources. The behavio