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Cryogenic Preservation and Storage of Animal Cells - Corning Life Science. PDF. - http://www.corning.com/Lifesciences/technical_information/techDocs/Cell_freezing_protocol.pdf

Category: Cell Biology and Cell Culture: Cell Culture Techniques: Cell Line Preservation Freezing

Cryogenic Preservation and Storage of Animal Cells. PDF.Corning Life Science. Very nice, detailed procedure for the freezing and storage of various cell lines. - [Read Cryogenic Preservation and Storage of Animal Cells - Corning Life Science. PDF.]

Cryogenic Preservation and Storage of Animal Cells Protocol - http://www.corning.com/Lifesciences/technical_information/techDocs/Cell_freezing_protocol.pdf

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

Cryogenic preservation (storage below -100°C) of cell cultures is widely used to maintain backups or reserves of cells without the associated effort and expense of feeding and caring for them. The success of the freezing process depends on four critical areas: Proper handling and gentle harvesting of the cultures; Correct use of the cryoprotective agent; A controlled rate of freezing; Storage under proper cryogenic conditions. - [Read Cryogenic Preservation and Storage of Animal Cells Protocol]

Culture Chambers for Live-Cell Imaging - http://www.microscopyu.com/articles/livecellimaging/culturechambers.html

Category: Microscopy Protocols: Live-Cell Imaging Protocols

Specimen chambers have had many designs published over the years describing systems that offer excellent optical properties while allowing specimens to be maintained for varying amounts of time. Ranging in complexity from the simple preparation of a sealed coverslip on a microscope slide to sophisticated perfusion chambers that enable tight control of virtually all environmental variables culture chambers are designed to to allow living specimens to be observed with minimal invasion at high res. - [Read Culture Chambers for Live-Cell Imaging]

Culture Conditions for Various Stable Cell Lines - http://www.flemingtonlab.com/Protocols/CultureStableCellLines.pdf

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

Culture conditions for various stable cell lines. - [Read Culture Conditions for Various Stable Cell Lines]

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

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

Culture of Human Embryonic Stem Cells (hESC). one simple protocol outlined. NIH Stem Cell Unit - [Read Culture of Human Embryonic Stem Cells (hESC)]

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 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]

Cytokine Bioassays Protocols - http://www.ebioscience.com/ebioscience/appls/BAC.htm?

Category: Signalling Signal Transduction Protocols: Cytokine Protocols

Cytokine-Induced Proliferation of Indicator Cell Lines, TNF-Î±-Induced Killing of L929 Cell Line, IFN-Î³ Protection from Viral Infection of Cell Lines. eBiosciences. - [Read Cytokine Bioassays Protocols]

Cytokine Neutralization, In Vitro Using Antibodies - http://www.ebioscience.com/ebioscience/appls/NU.htm

Category: Signalling Signal Transduction Protocols: Cytokine Protocols

Antibody Neutralization of TNF-Î±-Induced Killing of L929 Cell Line, IFN-Î³-Protection from Viral Infection of L929 and A549 Cell Lines, TNF-Î±-Induced Killing of L929 Cell Line. eBiosciences. - [Read Cytokine Neutralization, In Vitro Using Antibodies]

De Novo Isolation of Embryonic Stem (ES) Cell Lines from Blastocysts Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4403

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

The starting material for de novo isolation of stem cell lines can be either normal 3.5-days post coitum (dpc) expanded blastocysts or "delayed" blastocysts. Delayed blastocysts are usually collected 4-6 days after ovariectomy. For both groups of blastocysts, tissue culture procedures are similar. The only difference is the timing of the first disaggregation, because delayed blastocysts will initially grow more slowly. - [Read De Novo Isolation of Embryonic Stem (ES) Cell Lines from Blastocysts Protocol]

Dendritic Cell Derived-Exosomes - http://www.jleukbio.org/cgi/rapidpdf/jlb.0206094v1.pdf

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

Protocol for isolation of dendritic cell derived exosomes. - [Read Dendritic Cell Derived-Exosomes]

Derivation of Trophoblast Stem (TS) Cell Lines from Blastocysts Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4407

Category: Stem Cell Protocols

This protocol decribes derivation of TS cell lines from 3.5-days post coitum (dpc) mouse blastocysts. The procedure is similar to the derivation of embryonic stem (ES) cell lines. However, the success rate is considerably higher, and less expertise is required to recognize pluripotent TS cell colonies. - [Read Derivation of Trophoblast Stem (TS) Cell Lines from Blastocysts Protocol]

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

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Three Ambion kits were used to quantitate specific miRNAs and to detect differential miRNA expression in various mouse brain regions and cell types isolated by laser capture microdissection (LCM). These techniques can be applied to studying miRNA in other species, tissues, and cell types. Includes: Obtain Laser Capture Microdissected Samples; Isolate miRNA from LCM Samples; Quantitate miRNA by qRT-PCR. - [Read Detect and Quantitate MicroRNA in Laser Capture Microdissection Samples]

Detection of Fungi by Fluorescence Microscopy Using Fluorescent Brighteners - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./flbr.html~main

Category: Microscopy Protocols: Fluorescence Microscopy Protocols

Certain fluorescent dyes such as Blankophor have a high affinity for the b -glycosidically linked polysaccharides such as glucan and chitin, which are main the constituents of the fungal cell wall. Therefore, these fluorescent dyes can be used for screening clinical samples for the presence of fungal elements. This procedure can be performed using the following specimens: Nail, Skin, Bronchial alveolar lavage fluid, Sputum and Biopsies. - [Read Detection of Fungi by Fluorescence Microscopy Using Fluorescent Brighteners]

Detection of Mycoplasma in Mammalian Cell Cultures Using Fluorescence Microscopy Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4351

Category: Cell Culture Protocols: Cell Culture Contamination Prevention and Removal Protocols

Protocol describes a method for mycoplasma detection using Hoechst 33258, a fluorescent dye that stains DNA. Staining of particulate DNA matter around the cell nucleus (on the cell surface or in the cytoplasm) is indicative of mycoplasma contamination. - [Read Detection of Mycoplasma in Mammalian Cell Cultures Using Fluorescence Microscopy Protocol]

Determination of Neopterin and Biopterin by Liquid Chromatography Protocol - http://www.natureprotocols.com/2006/10/13/determination_of_neopterin_and.php

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol for determination of neopterin and biopterin by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) in rat and human plasma, cell extracts and tissue homogenates. - [Read Determination of Neopterin and Biopterin by Liquid Chromatography Protocol]

Determination of Tissue Fungal Burden Utilizing Quantitative Real Time Polymerase Chain Reaction - http://www.sacmm.org/pdf/Determination%20of%20Tissue%20Fungal%20Burden%20utilizing%20Quantitative%20Real%20Time%20PCR.pdf

Category: Fungi Protocols: Fungi Genetics Protocols

Standard operating procedure for the determination of tissue fungal burden utilizing quantitative real time polymerase chain reaction (QPCR). This standard operating procedure will provide information on how to assess fungal tissue burden of infected animals by use of a single copy (FKS) or multicopy gene (18s RNA) to assess the number of fungal cell nuclei present. - [Read Determination of Tissue Fungal Burden Utilizing Quantitative Real Time Polymerase Chain Reaction]

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]

Development of Techniques for Primary Culture of C. elegans Embryonic Neurons 2nd Part - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=15

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Development of techniques for primary culture of C. elegans embryonic neurons. Includes: Optimized cell culture protocol. - [Read Development of Techniques for Primary Culture of C. elegans Embryonic Neurons 2nd Part]

Differential Display-PCR Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3844

Category: PCR Protocols

Method uses PCR to amplify and display many cDNAs derived from the mRNAs of a given cell or tissue type. The method relies on two different types of synthetic oligonucleotides: anchored antisense primers and arbitrary sense primers. A typical anchored primer is complementary to approx. 13 nucleotides of the poly(A) tail of mRNA and the adjacent two nucleotides of the transcribed sequence. - [Read Differential Display-PCR Protocol]

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]

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]

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 28)

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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