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Neutral Red Cytotoxicity Assay Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/64.htm

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

The cytotoxic effect of chemicals upon mammalian cells, such as BALB/c 3T3 and HepG2, in culture is measured by highest tolerated dose (HTD), cell viability (Neutral Red) and total cell protein (coomassie blue). - [Read Neutral Red Cytotoxicity Assay Protocol]

Open Directory - Science: Biology: Botany: Tissue Culture - http://dmoz.org/Science/Biology/Botany/Tissue_Culture/

Category: Plant Biology Protocols: Plant Tissue Culture

Plant Tissue Culture - Laboratory protocol for micropropagation from stem tip, node, meristem, embryo or seed. Includes forum maintained by the National ... - [Read Open Directory - Science: Biology: Botany: Tissue Culture]

Optimized Protocols for Fluorescent in situ Hybridization in Drosophila Tissues - http://www.utoronto.ca/krause/documents/FISHCHAPTER.pdf

Category: Drosophila Protocols: Drosophila Genetics Protocols

Optimized protocols for fluorescent in situ hybridization in Drosophila tissues. Includes: RNA Probe Preparation; Initial Embryo Fixation; Post-Fixation, Hybridization and post-Hybridization Washes; Development of FISH Signal; Mounting and Viewing of Samples; Double FISH; FISH on Dissected Tissues; RNA-Protein Double-labeling. - [Read Optimized Protocols for Fluorescent in situ Hybridization in Drosophila Tissues]

Ovary Cell Proliferation Test - http://embryo.ib.amwaw.edu.pl/invittox/prot/36.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cell Specific Cytotoxicity Assays Protocols

The rate of cellular proliferation may be regarded as an overall indicator of the physiological status of the cell. Therefore, the effect of various toxic substances on different cell functions will be reflected by changes in the proliferation rate. - [Read Ovary Cell Proliferation Test]

Preparation and Fractionation of Drosophila Embryo Extracts for the In Vitro Characterization of RNA - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4324

Category: RNAi Technology Protocols: RNAi Drosophila Protocols

Protocol describes a method to prepare and fractionate Drosophila embryo extracts. These extracts can be used for the in vitro characterization of RNAi. - [Read Preparation and Fractionation of Drosophila Embryo Extracts for the In Vitro Characterization of RNA]

Preparation of siRNAs from Drosophila Embryo Extracts Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4325

Category: RNAi Technology Protocols: RNAi Drosophila Protocols

siRNAs produced upon the addition of dsRNA to Drosophila embryo extract are enriched in a micrococcus-nuclease-resistant fraction. After proteinase K treatment and dephosphorylation with calf intestinal phosphatase, these siRNAs mediate efficient RNAi in vitro. - [Read Preparation of siRNAs from Drosophila Embryo Extracts Protocol]

Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4400

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the preparation of feeder cells from MEF cells or from the STO mouse fibroblast cell line. The cells are rendered mitotically inactive by treatment with {gamma}-irradiation. The feeder layers can then be used to maintain embryonic stem (ES) cells in the undifferentiated state. - [Read Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells Protocol]

Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells: Treatment with Mitomyc - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4399

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the preparation of feeder cell layers from MEF cells or from the STO mouse fibroblast cell line. The cells are rendered mitotically inactive by treatment with mitomycin C. The feeder layers can then be used to maintain embryonic stem (ES) cells in the undifferentiated state. - [Read Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells: Treatment with Mitomyc]

Preparing Mouse Embryo Fibroblasts Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4398

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the preparation of mouse embryo fibroblasts (MEFs), which can then be used as feeder cells to maintain embryonic stem (ES) cells in the undifferentiated state. - [Read Preparing Mouse Embryo Fibroblasts Protocol]

Production of Completely ES Cell-Derived Fetuses by Aggregation with Tetraploid Embryos - http://www.mshri.on.ca/nagy/Tetraploid/tetra.htm

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Protocol for the production of completely ES cell-derived fetuses by aggregation with tetraploid embryos. Includes: Recovery of 2-cell stage embryos; Production of tetraploid embryos; Preparation of aggregation plate; Removal of Zona Pellucida; ES cells/ tetraploid embryo "SANDWICH" aggregation; Transfer of embryos. - [Read Production of Completely ES Cell-Derived Fetuses by Aggregation with Tetraploid Embryos]

Protocol to Count Cell Number of Preimplantation Embryos - http://www.animal.ufl.edu/hansen/protocols/embryocellnumber.htm

Category: Developmental Biology: Embryology Protocols

Protocol to Count Cell Number of Preimplantation Embryos using Nuclear Staining with Hoechst 33342 or DAPI. Includes: Preparation of Embryos; Preparation of Hoechst 33342 dye; Preparation of DAPI; Staining the Embryo; Mounting Embryos to Slides; What to Do When There are Too Many Cells to Count. - [Read Protocol to Count Cell Number of Preimplantation Embryos]

Rabbit Articular Chondrocyte Functional Toxicity Test - http://embryo.ib.amwaw.edu.pl/invittox/prot/41.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cell Specific Cytotoxicity Assays Protocols

In this test rabbit articular chondrocytes are cultured in the presence of test compound, the toxicity of which is then determined by its effect on the production of proteoglycan by the cells, as detected by the dye Alcian Blue. - [Read Rabbit Articular Chondrocyte Functional Toxicity Test]

Rat Hepatocyte Flow Cytometric Cytotoxicitiy Test - http://embryo.ib.amwaw.edu.pl/invittox/prot/23.htm#TOP

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

Flow cytometry is used to monitor drug-induced changes in DNA and protein contents of hepatocytes cultured at physiological oxygen concentrations. - [Read Rat Hepatocyte Flow Cytometric Cytotoxicitiy Test]

Red Blood Cell Test System Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/37.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cell Specific Cytotoxicity Assays Protocols

An in vitro red blood cell assay is presented which allows the estimation of the irritation potential of tensides and tenside containing materials such as shampoos, shower gels, cleaning products, etc. The estimation is based on the fact that surfactants interact strongly with cellular membranes and proteins. Both effects are measured photometrically by use of the inherent native dye, oxyhemoglobin. - [Read Red Blood Cell Test System Protocol]

RNA Whole Mount In Situ Hybridization Protocol - http://genetics.med.harvard.edu/~cepko/protocol/ctlab/ish.ct.htm

Category: Cytogenetics Protocols: In Situ Hybridization Tissue Sections Protocols

Protocol for RNA whole mount in situ hybridization. Includes: Embryo preparation; Prehybridization and Hybridization; Post-hybridization washes, blocking, and antibody incubation; Post-antibody washes; Color development. - [Read RNA Whole Mount In Situ Hybridization Protocol]

SIRC Cytotoxcitiy Test - http://embryo.ib.amwaw.edu.pl/invittox/prot/40.htm

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

Rabbit-derived corneal cells are cultured in the presence of test compounds, the toxicity of which are determined by their effect upon cell viability. A decrease in cell number, as measured by uptake of the dye Neutral Red, serves as an indicator of potential cytotoxicity. This test has been proposed as a potential replacement alternative for the Draize Eye Irritation test. - [Read SIRC Cytotoxcitiy Test]

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]

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]

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

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

V79 Cytotoxicity Test for Membrane Damage - http://embryo.ib.amwaw.edu.pl/invittox/prot/39.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

The cytotoxic effect of test chemicals in V79 cell culture can be determined by assessing damage to the plasma membrane as determined by a nucleic acid leakage assay. - [Read V79 Cytotoxicity Test for Membrane Damage]

Visualizing Early Embryo Implantation Sites by Dye Injection Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4361

Category: Developmental Biology: Embryology Protocols

This protocol describes a method for visualizing early embryo implantation sites using Chicago Sky Blue 6B dye. Once implantation and interimplantation sites are identified and separated, they can be used for cellular, biochemical, and molecular biology analyses. - [Read Visualizing Early Embryo Implantation Sites by Dye Injection Protocol]

Whole Mount In Situ Hybridization for the Detection of mRNA in Drosophila Embryos Protocol - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_208-215.pdf

Category: Drosophila Protocols: Drosophila Genetics Protocols

Protocol for whole mount in situ hybridization for the detection of mRNA in Drosophila embryos. Includes: Embryo collection and fixation; Prehybridization treatment; Hybridization; Washing and detection. - [Read Whole Mount In Situ Hybridization for the Detection of mRNA in Drosophila Embryos Protocol]

Articles (1-3 of 3)

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.

Picking Transfected ES Cell Clones Protocol

This protocol a protocol on how to generate transfected embryonic stem (ES) cell clones. The previous protocol in this series is the Protocol for Electroporation of ES cells. The next protocol in the series is the Protocol on Disaggregation, Expansion, and Freezing of Transfected ES Clones.

Vector Design Protocol for Transgenic Mice Generation

The protocol gives general considerations for the design of targeting vectors for transgenic mice. The protocol shares tips in the design of knock-out and knock-in vectors and some of their strategies for producing homologously recombined embryonic stem cells.

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