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Culture of Human Embryonic Stem Cells (hESC) Protocol - http://stemcells.nih.gov/research/NIHresearch/scunit/culture.asp

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Protocol for the culture of human embryonic stem cells (hESC). - [Read Culture of Human Embryonic Stem Cells (hESC) Protocol]

Culture of Human Prostatic Carcinoma Cell Lines - http://www.celldeath.de/apometh/rpmi.html

Category: Cell Biology and Cell Culture: Cell Line Culture Conditions

Culture of Human Prostatic Carcinoma Cell Lines, Growing and splitting the cells, Preparation of frozen stocks in liquid nitrogen, How to bring frozen cells back into culture, Concentrations of antibiotics for the selection of stable transfectants. LNCa - [Read Culture of Human Prostatic Carcinoma Cell Lines]

Culturing of Insect Cells and Production of Baculovirus Expressed Recombinant Proteins in 2.8 Liter - http://www.hyclone.com/pdf/procedure_insect_fernbach.pdf

Category: Cell Culture Protocols: Insect Cell Culture Protocols

The use of Ferbach flasks is an ideal method to amplify the cells and virus inoculum for the bioreactor. The method is also very practical for producing sufficient amounts of various proteins for feasibility studies using HyQ© SFX-Insect™ medium. The large surface area in this vessel enables sufficient aeration and, therefore, the cells, when they reach high density, are not oxygen depleted. - [Read Culturing of Insect Cells and Production of Baculovirus Expressed Recombinant Proteins in 2.8 Liter]

Culturing Schwann Cells from Adult Peripheral Nerve Protocol - http://www.physiol.usyd.edu.au/daved/papers/1998/sc_culture/

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols: Rat Neuronal Cell Culture Protocols

Protocol for culturing schwann cells from adult peripheral nerve. - [Read Culturing Schwann Cells from Adult Peripheral Nerve Protocol]

Culturing Trophoblast Stem (TS) Cell Lines Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4406

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Culture conditions have been established for a second blastocyst-derived cell line, trophoblast stem (TS) cells, in addition to embryonic stem (ES) cells. This protocol describes a method for culturing TS cell lines. These cells can then be used to study trophoblast differentiation and placental function. - [Read Culturing Trophoblast Stem (TS) Cell Lines Protocol]

CYP1A1-Inducing Potency and Cytotoxicity Test in the HEPA-1 Mouse Hepatoma Cell Line - http://embryo.ib.amwaw.edu.pl/invittox/prot/112.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Liver Toxicity Assays Protocols

This bioassay utilizes cultured Hepa-lclc7 (Hepa-1) mouse hepatoma cells to assess the CYPlA1-inducing potency or cytotoxicity of pure test chemicals or environmental samples. In the Hepa-l induction test , the CYPlA1-inducing potency of the test sample is detected as increased aryl hydrocarbon hydroxylase (AHH) and 7-ethoxyresorufin O-deethylase (EROD) activities. - [Read CYP1A1-Inducing Potency and Cytotoxicity Test in the HEPA-1 Mouse Hepatoma Cell Line]

Defrosting Eukaryotic Cells Frozen In Liquid N2 Protocol - http://www2.umdnj.edu/~geller/lab/thawing.html

Category: Cell Culture Protocols: Cell Storing and Cell Freezing Protocols

Protocol for the defrosting of eukaryotic cells frozen in liquid N2. - [Read Defrosting Eukaryotic Cells Frozen In Liquid N2 Protocol]

Denaturing Protein Immunoprecipitation from Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4619

Category: Immunology: Immunoprecipitation Protocols: Protein Immunoprecipitation Protocols

Protocol describes a denaturing immunoprecipitation (IP) for mammalian cells. Prefer to use denaturing IPs to recover labeled proteins from pulse-chase experiments. The use of denaturing IPs reduces background considerably. - [Read Denaturing Protein Immunoprecipitation from Mammalian Cells Protocol]

Detect and Quantitate MicroRNA in Laser Capture Microdissection Samples - http://www.ambion.com/techlib/tn/131/6.html

Category: RNAi Technology Protocols

Information on how detect and quantitate MicroRNA in laser capture microdissection samples. Includes: Obtain Laser Capture Microdissected Samples; Isolate miRNA from LCM Samples; Quantitate miRNA by qRT-PCR; Detection of miRNA in Microdissected Tissue from Mouse Brain by qRT-PCR; Differential Expression of MicroRNA in Whole Brain Tissue Compared to a More Homogeneous Population of Cells. - [Read Detect and Quantitate MicroRNA in Laser Capture Microdissection Samples]

Detection of BrdU Incorporation in DNA Synthesizing Cells Protocol - http://www.bdbiosciences.com/pharmingen/protocols/BrdU_Incorporation.shtml

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Apoptosis Cytometry Protocols

Protocol for the detection of BrdU incorporation in DNA synthesizing cells. - [Read Detection of BrdU Incorporation in DNA Synthesizing Cells Protocol]

Detection of Chromosomal Imbalances Using DOP-PCR and Comparative Genomic Hybridization (CGH) - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_95-103.pdf

Category: Cytogenetics Protocols

CGH provides a comprehensive picture of all chromosomal gains and losses within a single experiment. In contrast to banding analysis, CGH requires no preparation of metaphase chromosomes from the cells to be analyzed (which in certain tumor types may be difficult or even impossible). - [Read Detection of Chromosomal Imbalances Using DOP-PCR and Comparative Genomic Hybridization (CGH)]

Detection of Protein Phosphorylation in Tissues and Cells Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&objectId=6677CD53C088A0E7A7B22F422FFCDC12&treeNodeId=9E6639B7FED0C828BBFE860A8E957694

Category: Immunology: Immunoprecipitation Protocols: Immunoprecipitation of Radiolabeled Cells Protocols

Most powerful and convincing method to determine if a specific protein is phosphorylated in a physiologically relevant manner is to assay phosphorylation in situ. The procedure described involves incubating cultured cells (e.g., primary neuronal cultures or transfected cells) or tissue preparations (e.g., hippocampal slices) with [32P]orthophosphate, which is then taken up by the cells or tissues and incorporated into the γ-phosphate position of ATP. - [Read Detection of Protein Phosphorylation in Tissues and Cells Protocol]

Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4345

Category: RNAi Technology Protocols: RNAi Mammalian Cells Protocols

Protocol describes the use of western blotting to analyze the RNAi-induced depletion of target protein from mammalian cells. Immunofluorescence can be used as an alternative. - [Read Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting Protocol]

Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4345

Category: Western Blot Protocols

Determining Cell Cycle Stages by Flow Cytometry Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E662F660A11AB3ABD97E38B90B2AF0D&objectid=6674031CC5AE0F442766156EA393FE81

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Cycle Analysis Protocols

Describes assays used to determine the distribution of a population of cells to the different stages of the cell cycle as analyzed by flow cytometry. Staining the DNA with different fluorescent dyes, propidium iodide or DAPI, is one of the most direct ways of staging the cells based on DNA content. - [Read Determining Cell Cycle Stages by Flow Cytometry Protocol]

Determining Number of Live and Dead Cells in Cell Population: Cytotoxicity Assay Protocol - http://www.promega.com/paguide/chap4.htm#title4

Category: Cell Biology and Cell Culture: Cell Viability Protocols

Cell-based assays are important tools for contemporary biology and drug discovery because of their predictive potential for in vivo applications.This assay gives ratiometric, inversely proportional values of viability and cytotoxicity (Figure 4.15) that are useful for normalizing data to cell number. Also, this reagent is compatible with additional fluorescent and luminescent chemistries. - [Read Determining Number of Live and Dead Cells in Cell Population: Cytotoxicity Assay Protocol]

Determining Plating Efficiency in Yeast: Direct Method - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4185

Category: Yeast Cell Culture Protocols: Yeast Sporulation Protocols

Direct method for determining efficiency in yeast. The plating efficiency of a strain is a measure of the percentage of cells in a culture that are capable of forming colonies (colony forming . - [Read Determining Plating Efficiency in Yeast: Direct Method]

Differentiate ES cells into cardiac myocytes - http://www.mshri.on.ca/nagy/Protocols/cardiac.htm

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

Differentiate ES cells into cardiac myocytes. Nagy Lab - [Read Differentiate ES cells into cardiac myocytes]

Differentiate ES cells into cystic embryoid bodies - http://www.mshri.on.ca/nagy/Protocols/cystemb.htm

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

Differentiate ES cells into cystic embryoid bodies. Nagy Lab - [Read Differentiate ES cells into cystic embryoid bodies]

Differentiate ES cells into glial cells and neurons - http://www.mshri.on.ca/nagy/Protocols/neurdif.htm

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

Differentiate ES cells into glial cells and neurons. Nagy Lab. - [Read Differentiate ES cells into glial cells and neurons]

Differentiating Embryonic Stem (ES) Cells into Embryoid Bodies Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4405

Category: Stem Cell Protocols

Pluripotent ES cells can develop into many types of differentiated tissues if they are placed back into a differentiating environment. Often, differentiation proceeds through an intermediate stage called the embryoid body (EB). EBs can be manipulated further to generate more differentiated cell types. This protocol describes a method for differentiation of ES cells into EBs. - [Read Differentiating Embryonic Stem (ES) Cells into Embryoid Bodies Protocol]

Differentiation of Embryonic Stem (ES) Cells Using the Hanging Drop Method - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4485

Category: Stem Cell Protocols: Stem Cell Culture Protocols

In vitro differentiation of ES cells occurs when the cells are allowed to aggregate in suspension culture in the absence of mouse embryonic fibroblast (MEF) feeders and leukemia inhibitory factor (LIF). Hanging drops provide a uniform aggregate size, which is then expanded by continued growth in suspension culture. The embryoid bodies are then plated and allowed to differentiate further in culture. - [Read Differentiation of Embryonic Stem (ES) Cells Using the Hanging Drop Method]

Direct Adaptation of Insect Cells to HyQ© SFX-Insect™ Medium - http://www.hyclone.com/pdf/procedure_insect_directadaptation.pdf

Category: Cell Culture Protocols: Insect Cell Culture Protocols

Using this method, insect cells should be adapted to HyQ SFX-Insect in 4-8 passages. However, there are some cell lines and clones that are more sensitive to the physiochemical and nutritional changes, and in some cases it may take longer than 8 passages for the adaptation. - [Read Direct Adaptation of Insect Cells to HyQ© SFX-Insect™ Medium]

Direct PCR from Whole Yeast Cells Protocol - http://www.duke.edu/web/ceramide/protocols/0003.html

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast PCR Protocols

Protocol for direct PCR from whole Yeast cells. - [Read Direct PCR from Whole Yeast Cells Protocol]

Disaggregating Cleavage-Stage Embryos and the Inner Cell Mass of Blastocysts into Individual Cells - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4427

Category: Mouse Protocols: Mouse Embryology Protocols

Protocol describes a method for disaggregating embryos and the ICM of blastocysts into individual cells. - [Read Disaggregating Cleavage-Stage Embryos and the Inner Cell Mass of Blastocysts into Individual Cells]

Articles (1-10 of 24)

Lambda Phage Protocol Large DNA Preparation

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

Genomic DNA Labeling of Microarrays Protocol

DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.

Drosophila Cell Transfection Protocol

A simple Drosophila tissue culture cell transfection method.

Tubulin Polymerization Protocol using GTP and GMPCPP

Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.

Acid Guanidinium Thiocyanate RNA Isolation Phenol Chloroform Extraction

A single step RNA isolation protocol using Phenol Chloroform Extraction and Acid Guanidinium Thiocyanate. This RNA isolation method uses the fact that guanidinium thiocyanate can simultaneously lyse the cells and inactive cellular RNAses during the initial RNA isolation step allow a single step in the method.

Microinjection Pipettes Protocol

Microinjection Pipettes Preparation Protocol.

Acid Washing Microinjection Pipettes Protocol

Acid Washing of Glass microinjection pipettes protocol.

Paraffin Embedding Protocol

Paraffin Embedding Protocol for molecular profiling. This Paraffin Embedding Protocol describes the processing of the tissues into sections following ethanol fixation. Molecular profiling (MP) is a technique that is used to visualize the global patterns of RNA expression or protein expression in various cell types and disease processes.

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