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Simple and Reliable Method To Precipitate Proteins from Bacterial Culture Supernatant PDF - http://aem.asm.org/cgi/reprint/70/1/610.pdf

Category: Protein Protocols: Protein Precipitation Procols

Simple and Reliable Method To Precipitate Proteins from Bacterial Culture Supernatant. Randolph B. Caldwell et al., AEM ASM. - [Read Simple and Reliable Method To Precipitate Proteins from Bacterial Culture Supernatant PDF]

Simplifying hESC culture - http://www.bloodjournal.org/cgi/content/full/105/12/4550

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

Human embryonic stem cells are a valuable resource for research and cell replacement therapy but are notoriously cumbersome to culture. Bhatia and colleagues show that an increased dose of basic fibroblast growth factor eliminates the need for feeder laye - [Read Simplifying hESC culture]

Single Strand DNA Prep for Sequencing Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/S2.html

Category: DNA Protocols: DNA Sequencing Protocols

Protocol for single strand DNA prep for sequencing. Protocol works well from a 5 ml starting culture or a 1 ml starting culture. - [Read Single Strand DNA Prep for Sequencing Protocol]

Single Strand DNA Prep For Sequencing Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/S2.html

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols

Protocol for single strand DNA prep for sequencing. This protocol works well from a 5 ml starting culture or a 1 ml starting culture (adjust volume accordingly). - [Read Single Strand DNA Prep For Sequencing Protocol]

Sorting Transfected Cells Based on Gene Expression, Followed by Culture in Vivo - http://www.bdbiosciences.com/pdfs/whitePapers/23-2787-02.pdf

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

Describes how FACSort can be used to enrich for transfected mouse cells expressing high levels of the human thrombin receptor. The sorted fraction can then be cultured in vivo and reanalyzed 12 days later to show that it remains enriched for thrombin receptor-expressing cells. - [Read Sorting Transfected Cells Based on Gene Expression, Followed by Culture in Vivo]

Split Luciferase Complementation Assay for Studying Interaction of Proteins X and Y in Living Mice - http://www.cshprotocols.org/cgi/content/abstract/2006/29/pdb.prot4595

Category: Reporter Gene Assays: Luciferase Assay Protocols

Protocol describes a split luciferase complementation assay that can be used to repetitively and noninvasively study the interaction of proteins in small living animals. After the expression of the appropriate vectors has been checked in cell culture in vivo, studies can be performed either by implanting transiently transfected cells for short-term analysis (maximum of 7 days), or with tumor models grown from tumor cells stably expressing the complete reporter system. - [Read Split Luciferase Complementation Assay for Studying Interaction of Proteins X and Y in Living Mice]

Stable Isotope Labeling by Amino Acids in Cell Culture SILAC Protocol - http://www.natureprotocols.com/2007/01/11/a_practical_recipe_for_stable.php

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol for stable isotope labeling by amino acids in cell culture (SILAC). Protocol describes how to apply SILAC and the use of nano-scale liquid chromatography coupled to electrospray ionization mass spectrometry for protein identification and quantification. This procedure can be completed in 8 days. - [Read Stable Isotope Labeling by Amino Acids in Cell Culture SILAC Protocol]

Static Culture of Postimplantation Embryos for Imaging Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4459

Category: Primary Cell Isolation and Culture Protocols

This protocol describes a method for static culture of early postimplantation mouse embryos on a microscope stage. Embryos between 6.5 and 9.5 days post coitum (dpc) can be cultured and imaged for 24 hours, with very little growth retardation. - [Read Static Culture of Postimplantation Embryos for Imaging Protocol]

Static Culture of Postimplantation Embryos for Imaging Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4459

Category: Mouse Protocols: Mouse Cell Culture Protocols

Protocol describes a method for static culture of early postimplantation mouse embryos on a microscope stage. Embryos between 6.5 and 9.5 days post coitum (dpc) can be cultured and imaged for 24 hours, with very little growth retardation. - [Read Static Culture of Postimplantation Embryos for Imaging Protocol]

Static Culture of Postimplantation Embryos Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4373

Category: Cell Culture Protocols

Protocol describes static culture of postimplantation embryos, an alternative to the roller method. The static method is best suited to 6.0 to 7.0 days post coitum (dpc) embryos followed for 24 hours (7.0 dpc embryos) to 48 hours (6.0 dpc embryos) of development. It allows repetitive real-time observation with minimal handling of the embryo. It is especially useful if single or small groups of embryos need to be distinguished from each other. - [Read Static Culture of Postimplantation Embryos Protocol]

Sterilization and Filtration Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E662B9D0482A943EC97AC4807854493&objectid=6673A220D50910E56F81295AE515690F

Category: Cell Culture Protocols: Sterile Technique Protocols

This unit on sterility in the tissue culture environment describes methods for sterilization of liquid and dry goods used for tissue culture and filtration of liquids to prevent contamination of cultures. - [Read Sterilization and Filtration Protocol]

Sterilization of Plant Culture Media Protocol - http://www.sigmaaldrich.com/Area_of_Interest/Life_Science/Plant_Biotechnology/Tissue_Culture_Protocols/Media_Sterilization.html

Category: Cell Biology and Cell Culture: Cell Culture Growth Media: Plant Cell Culture Growth Media Protocols

Protocol for sterilization of plant culture media. Includes: Minimum Autoclaving Time for Plant Tissue Culture Medium. - [Read Sterilization of Plant Culture Media Protocol]

Storage of Tissue Culture Supernatants Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4280

Category: Antibody Protocols: Antibody Handling Protocols

This protocol describes the methods of storage for hybridoma tissue-culture supernatants. - [Read Storage of Tissue Culture Supernatants Protocol]

Studying Mitosis in Cultured Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4674

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

Protocol describes methods for maintaining healthy, dividing mammalian cells in culture and during imaging, when mitosis can be examined. Rose chambers are preferable for observation and microinjection of living mitotic cells, but slide/coverslip preparations are easy to make and do not require any special equipment. - [Read Studying Mitosis in Cultured Mammalian Cells Protocol]

Studying Mitosis in Cultured Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4674

Category: Cytogenetics Protocols

Protocol describes methods for maintaining healthy, dividing mammalian cells in culture and during imaging, when mitosis can be examined. Rose chambers are preferable for observation and microinjection of living mitotic cells, but slide/coverslip preparations are easy to make and do not require any special equipment. Another inexpensive and easy-to-use alternative is to grow cells in a culture dish with a glass bottom. Such dishes are suitable for microinjection experiments. - [Read Studying Mitosis in Cultured Mammalian Cells Protocol]

Studying Mitosis in Cultured Mammalian Cells Prtocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4674

Category: Cell Biology and Cell Culture: Cell Cycle Protocols: Mitosis Protocols

Protocol describes methods for maintaining healthy, dividing mammalian cells in culture and during imaging, when mitosis can be examined. Rose chambers are preferable for observation and microinjection of living mitotic cells, but slide/coverslip preparations are easy to make and do not require any special equipment. Another inexpensive and easy-to-use alternative is to grow cells in a culture dish with a glass bottom. Such dishes are suitable for microinjection experiments. - [Read Studying Mitosis in Cultured Mammalian Cells Prtocol]

Synchronization of Mammalian Cell Cultures in Mitosis Using Selective Detachment Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4489

Category: Cell Biology and Cell Culture: Cell Cycle Protocols: Mitosis Protocols

Protocol describes a method for synchronizing monolayer cells in mitosis using selective detachment from their substrate. During mitosis, cells become spherical, causing them to become more loosely attached to their substrate. The "rounded up" cells are selectively detached by tapping the culture flask, resulting in a population in which as many as 90-98% of the cells are in mitosis. The drug nocodazole is used to increase the percentage of cells undergoing mitosis before detachment is performed - [Read Synchronization of Mammalian Cell Cultures in Mitosis Using Selective Detachment Protocol]

Synchronization of Mammalian Cell Cultures in Mitosis Using Selective Detachment Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4489

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

This protocol describes a method for synchronizing monolayer cells in mitosis using selective detachment from their substrate. During mitosis, cells become more spherical, causing them to become more loosely attached to their substrate. The "rounded up" cells are selectively detached by tapping the culture flask, resulting in a population in which as many as 90-98% of the cells are in mitosis. The drug nocodazole is used to increase the percentage of cells undergoing mitosis before detachment.. - [Read Synchronization of Mammalian Cell Cultures in Mitosis Using Selective Detachment Protocol]

Techniques for Primary Culture of C. elegans Embryonic Neurons - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=14

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

Development of techniques for primary culture of C. elegans embryonic neurons. - [Read Techniques for Primary Culture of C. elegans Embryonic Neurons]

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

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

Development of techniques for primary culture of C.elegans embryonic neurons. - [Read Techniques for Primary Culture of C. elegans Embryonic Neurons II]

Techniques for Primary Culture of C. elegans Embryonic Neurons III - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=16

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

Development of techniques for primary culture of C. elegans embryonic neurons - [Read Techniques for Primary Culture of C. elegans Embryonic Neurons III]

Tetrahymena Thermophila Ocular Irritancy Test - http://embryo.ib.amwaw.edu.pl/invittox/prot/22.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Microorganism Cytotoxicity Assays Protocols

The basis of this test is that a cytotoxic chemical (regardless of site or mechanism of action) will interfere with the normal motility of the protozoan, Tetrahymena thermophila, in culture. The degree of interference of motility as compared to control cultures, related to the concentration of the test compound, provides an indication of toxicity. - [Read Tetrahymena Thermophila Ocular Irritancy Test]

The Frame Cytotoxicity Test Kenacid Blue - http://embryo.ib.amwaw.edu.pl/invittox/prot/15.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 the change in total cell protein arising from the inhibition of cell proliferation (Kenacid Blue R dye binding method). - [Read The Frame Cytotoxicity Test Kenacid Blue]

The Isolation and Culture of Rat Hepatic Cells Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/7.htm

Category: Primary Cell Isolation and Culture Protocols

This method enables the isolation of different populations of liver cells and describes their subsequent culture. - [Read The Isolation and Culture of Rat Hepatic Cells Protocol]

The Isolation and Culture of Rat Hepatic Cells Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/7.htm

Category: Mouse Protocols: Mouse Cell Culture Protocols

The liver of a rat is cannulated and perfused in situ with buffer, following which it is excised and perfused in a closed system with a collagenase solution. After a period of time the liver begins to break up, at which point it is transferred to a measuring cylinder and culture medium is added. It is then gently agitated to cause the release of cells which are subsequently filtered and allowed to settle out. The parenchymal and non-parenchymal cells form two distinct layers which can be separat - [Read The Isolation and Culture of Rat Hepatic Cells 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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