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ES mouse cell stem cell Culture - http://imgen.bcm.tmc.edu/dtmc/mouse_embryonic_stem_cell_culture.html

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

Mouse Embryonic Stem Cell Culture. Southern Analysis of ES cells. The Darwin Transgenic Core Facility - [Read ES mouse cell stem cell Culture]

Feeder Cell Density - A key Parameter in Human Embryonic Stem Cell Culture - http://www.bioone.org/bioone/?request=get-document&issn=1543-706X&volume=040&issue=08&page=0255

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: ES Cell Growth Media and Requirements

Feeder Cell Density - A key Parameter in Human Embryonic Stem Cell Culture - [Read Feeder Cell Density - A key Parameter in Human Embryonic Stem Cell Culture]

FIXATION PROTOCOL FOR FLOW CYTOMETRIC ANALYSIS OF DNA BY PI STAINING - http://www.vetmed.auburn.edu/index.pl/cell_cycle/dna_content

Category: Histology Protocols: Fixation of Cells Tissues Protocol

Analysis of Cell Cycle Based on DNA Content. FIXATION PROTOCOL FOR FLOW CYTOMETRIC ... Liberate cells from tissue culture flask by trypsin digestion and ... FIXATION PROTOCOL FOR FLOW CYTOMETRIC ANALYSIS OF DNA BY PI STAINING. Auburn University, College of Veterinary Medicine. You. - [Read FIXATION PROTOCOL FOR FLOW CYTOMETRIC ANALYSIS OF DNA BY PI STAINING]

Fixing Attached Cells in Organic Solvents Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4293

Category: Cell Culture Protocols: Cell Fixing Protocols

Organic solvents such as alcohols and acetone remove lipids and dehydrate cells, precipitating the proteins on the cellular architecture. Be aware that different antigens may be affected differently by the various solvents. If no previous data are available for your antigen, start with the 50/50 mixture. For tissue culture dishes, concentrations of acetone higher than 50% will destroy the integrity of the plastic. - [Read Fixing Attached Cells in Organic Solvents Protocol]

Formaldehyde Treatment of Tissue Culture Hoods - http://www.nottingham.ac.uk/~mbzspd/methods/Formaldehyde_Treatme.html

Category: General Research Protocols and Methods: Sterile Technique

Formaldehyde sterilization of tissue culture hoods. A bit toxic however effective. Dr.Dawson, Department Biochemistry, Univ. Nottingham Medical School. - [Read Formaldehyde Treatment of Tissue Culture Hoods]

Frame Modified Neutral Red Uptake Cytotoxcity Test - http://embryo.ib.amwaw.edu.pl/invittox/prot/3.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

The cytotoxic effect of chemicals upon cells in culture is measured by cell viability (neutral red uptake) method. - [Read Frame Modified Neutral Red Uptake Cytotoxcity Test]

Freezing and Thawing of Embryonic Stem (ES) Cells Using Cryovials Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4402

Category: Stem Cell Protocols

This protocol describes a method for freezing and thawing ES cells using cryovials. It is important to freeze ES cell stocks as soon as possible to reduce the time that they are in culture. A careful record should be kept of the number of times cells are passaged and the location of the cryovials. - [Read Freezing and Thawing of Embryonic Stem (ES) Cells Using Cryovials Protocol]

Freezing and Thawing of Mammalian Cell Lines - http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Immunology/CellFreeze.html

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

Freezing and Thawing of Mammalian Cell Lines. For long term storage of myeloma cells, hybridoma cells, T cells, and other mammalian cell lines in liquid nitrogen, and restoring them in culture. Kitto Lab Texas. - [Read Freezing and Thawing of Mammalian Cell Lines]

Freezing and Thawing of Mammalian Cell Lines Protocol - http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Immunology/CellFreeze.html

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

For long term storage of myeloma cells, hybridoma cells, T cells, and other mammalian cell lines in liquid nitrogen, and restoring them in culture. - [Read Freezing and Thawing of Mammalian Cell Lines Protocol]

Fundamental Techniques in Cell Culture: A Handbook - http://www.sigmaaldrich.com/Area_of_Interest/Life_Science/Cell_Culture/Key_Resources/ECACC_Handbook.html

Category: Cell Biology and Cell Culture

Fundamental Techniques in Cell Culture: A Handbook. Excellent Resource. Sigma. - [Read Fundamental Techniques in Cell Culture: A Handbook]

G1 Synchronization of CHO Cells by Isoleucine Deprivation Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4486

Category: Cell Biology and Cell Culture: Cell Cycle Protocols: G Phase Protocols

This protocol provides a method for the synchronization of a monolayer culture of CHO cells in G1 using isoleucine deprivation. Since CHO cells can also be adapted to grow in suspension culture, this procedure can be used to obtain larger quantities of cells. When isoleucine is replaced, the cells resume growth and begin to enter S phase ~4 hours later. This method arrests almost 100% of the CHO cells in G1, and upon reversal, leads to rapid recovery of cell growth and very high cell viability. - [Read G1 Synchronization of CHO Cells by Isoleucine Deprivation Protocol]

G1 Synchronization of CHO Cells by Isoleucine Deprivation Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4486

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

General Cell Culture - http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Immunology/GeneralCulture.html

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

Covers general things you need to know to get started in cell culture work. Includes: Sterile Technique; Growth of Your Cell Culture; - [Read General Cell Culture]

General Method for Ras, Rac, Cdc42, and Rho Activation Assay - http://www.upstate.com/misc/protocol_detail.q.prot.e.general_method_ras.a.name.e.General_Method_for_Ras_Rac_Cdc42_and_Rho_Activation_Assay

Category: Signalling Signal Transduction Protocols

General protocol for Ras, Rac, Cdc42, and Rho activation assay. Includes: Affinity Precipitation/Immunoblot Protocol, Cell Culture and Extract Preparation (Adherent and Non Adherent cells), GTPÎ³S/GDP Loading for Positive and Negative Controls, Ras, Rac ,Cdc42, and Rho Pull-Down Assay and Western Blot and Detection. - [Read General Method for Ras, Rac, Cdc42, and Rho Activation Assay]

Germ Layer Explant Recombination Culture Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4369

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

This protocol describes a method for recombining and culturing germ layer fragments. It is useful for testing the inductive properties of fragments from wild-type and mutant mouse embryos. - [Read Germ Layer Explant Recombination Culture Protocol]

Germ Layer Explant Recombination Culture Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4369

Category: Mouse Protocols: Mouse Cell Culture Protocols

Protocol describes a method for recombining and culturing germ layer fragments. It is useful for testing the inductive properties of fragments from wild-type and mutant mouse embryos. - [Read Germ Layer Explant Recombination Culture Protocol]

Growing Adherent Cells Directly on Tissue Dishes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4290

Category: Cell Culture Protocols

For low-resolution work, cells to be used for staining can be grown directly on regular tissue-culture dishes. It is a convenient method that does not require much preparatory work. - [Read Growing Adherent Cells Directly on Tissue Dishes Protocol]

Growing Adherent Cells on Coverslips or Multiwell Slides Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4289

Category: Cell Culture Protocols

Glass is an excellent substrate for most tissue-culture-adapted cells and is compatible with all fixing and staining solutions. Glass coverslips in tissue-culture dishes or in 24-well multiwell plates are suitable carriers, as are multiwell slides. For high-resolution studies, choose glass coverslips of the highest available grade; #1 or #1.5 coverslips are the appropriate thickness. - [Read Growing Adherent Cells on Coverslips or Multiwell Slides Protocol]

Growing Bacteriophage M13 in Liquid Culture Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3994

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

Most manipulations with M13, including preparations of viral stocks and isolation of single- and double-stranded DNAs, begin with small-scale liquid cultures that are infected with an M13 plaque, picked from an agar plate. - [Read Growing Bacteriophage M13 in Liquid Culture Protocol]

Growth of Primary Cell Culture and Viral Vector Handling Protocols - http://www.stanford.edu/dept/EHS/prod/researchlab/bio/docs/Palmer_Bio_protocol.pdf

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

The procedures involve the isolation and growth of primary cell cultures from rodent and human tissue as well as the use of viral vectors for the introduction and expression of mammalian genes in cells in culture and in live rodents. - [Read Growth of Primary Cell Culture and Viral Vector Handling Protocols]

Guide For Identifying And Correcting Common Cell Growth Problems - http://www.corning.com/Lifesciences/technical_information/techDocs/cc_guide_identifing_correcting_common_cell_growth_problem_5_2_03_cls_cc_013w.pdf

Category: Cell Biology and Cell Culture: Troubleshooting Cell Culture Problems

Guide For Identifying And Correcting Common Cell Growth Problems. Corning. Surface Treatment Process, Problems Related to Technique, Problems Related to Incubators, Problems Related to Culture Media, Problem Solving Suggestions. - [Read Guide For Identifying And Correcting Common Cell Growth Problems]

Handbook of Cell Culture. - Sigma Aldrich. - http://www.sigmaaldrich.com/Area_of_Interest/Life_Science/Cell_Culture/Key_Resources/ECACC_Handbook/Cell_Culture_Techniques_12.html

Category: Cell Biology and Cell Culture: Cell Culture Techniques

ECACC Handbook of Cell Culture. Fundamental Techniques in Cell Culture: A Labratory Handbook. Sigma Aldrich. Excellent Guide to Cell Culture, aseptic technique, storage of cell lines, subculture of cells. - [Read Handbook of Cell Culture. - Sigma Aldrich.]

Hippocampal Neuron Cell Cultures - http://www.pharm.stonybrook.edu/faculty/tsirka/lab/HippocampalNeuronCultures.doc

Category: Neuroscience: Neuron Cell Culture Protocols

Hippocampal Neuron Culture Protocol, methods. Jean Siao, Ph.D. Tsirka Lab. - [Read Hippocampal Neuron Cell Cultures]

Histone H1 Kinase Activity Assay - http://www.bio.com/protocolstools/protocol.jhtml?id=p910

Category: Signalling Signal Transduction Protocols

This protocol describes assaying kinase activity of a putative kinase using Histone H1 as the substrate. Histone H1 is the canonical kinase substrate in this type of assay. Phosphorylation of Histone H1 is assessed by SDS-Polyacrylamide gel electrophoresis followed by autoradiography. Includes information: H1 Kinase Assay on Individual Xenopus Oocytes; H1 Kinase Assay on Xenopus Egg Extract Samples; H1 Kinase Assay on Tissue Culture Cells; Helpful protocol hints. - [Read Histone H1 Kinase Activity Assay]

Homogeneous Caspase 3-7 Assay Protocol - http://www.promega.com/tbs/tb295/tb295.pdf

Category: Pharmacology and Toxicology Protocols: Enzyme Assays Protocols

Homogeneous caspase-3/7 assay. Includes: Detection of Caspase-3/7 Activity in Cell Culture; Detection of Caspase-3 or -7 Activity in Purified Caspase Preparations; Positive and Negative Cell Culture Controls. - [Read Homogeneous Caspase 3-7 Assay Protocol]

Articles (1-7 of 7)

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.

Drosophila Cell Transfection Protocol

A simple Drosophila tissue culture cell transfection method.

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.

Immobilized Antigen Phage Antibody Selection Protocol

A protocol for the selection of Phage Antibodies using Immobilized Antigen. This method describes the selection of antibodies from bacteriophage antibody libraries that recognize a specific antigen. The phage display library of antibody-displaying phage particles is exposed to antigen attached to a solid substrate (Nunc Immuno™ tubes). The phage particles with affinity for antigen bind to the immobilized antigen and are selected from the library of phage expressing antibodies.

Histone H1 Kinase Activity Assay

Histone H1 Kinase Activity Assay Protocol. This protocol describes assaying kinase activity of a putative kinase using Histone H1 as the substrate. Histone H1 is the canonical kinase substrate in this type of assay. Phosphorylation of Histone H1 is assessed by SDS-Polyacrylamide gel electrophoresis followed by autoradiography.

Electrotransformation of BMH 81-17mut S for Isolating Site-Directed dsDNA Mutants

This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.

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