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Hamster Ovary Cell NA+/K+ -ATPase Test - http://embryo.ib.amwaw.edu.pl/invittox/prot/35.htm

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

Accumulation of lipophilic substances in the plasma membrane may affect the membrane lipid order and consequently affect the function of these proteins. Changes in the activity of the Na+/K+ -ATPase, which is the major active transport system responsible for the electrochemical potential in mammalian cells, can therefore be an indication of the effect that a chemical may have on the viability of the cell membrane and possibly the whole cell. - [Read Hamster Ovary Cell NA+/K+ -ATPase Test]

Hepatoma Cell Cultures as In Vitro Models for Hepatotoxicity - http://embryo.ib.amwaw.edu.pl/invittox/prot/13.htm

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

This test is designed to detect irreversible toxic effects on both cell growth and survival, by the evaluation of colony-forming (CF) efficiency, in hepatoma cell lines derived from man, rat and mouse. - [Read Hepatoma Cell Cultures as In Vitro Models for Hepatotoxicity]

Human Embryonic Stem Cell Protocols - http://stemcells.nih.gov/staticresources/research/protocols/BresaGen_hESC_manual_2.1.pdf

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Human embryonic stem cells (hESCs) are pluripotent cells capable of differentiation to representatives of all three germ layers. Includes: Isolation of Primary Mouse Embryo Fibroblasts; Thawing and preparing p1 MEF feeder plates; Preparation of MEF- Conditioned Medium (MEF-CM; Microdissection Passaging of hESCs; Bulk passaging of hESC; Cryopreservation of hESCs; Thawing of hESCs; Karyotyping. - [Read Human Embryonic Stem Cell Protocols]

Human Lymphocyte Cytotoxicity Assay Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/6.htm

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

This method measures the leakage of DNA and lactate dehydrogenase from lymphocytes into the surrounding medium as an indicator of cytotoxicity. This method also includes an assay of intracellular (mitochondrial) diaphorase as a measure of cellular activity (MTT assay). - [Read Human Lymphocyte Cytotoxicity Assay Protocol]

Human Oesophageal Culture Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/28.htm#TOP

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

This procedure describes a method for establishing short-term explant cultures of oesophageal mucosa. Adverse effects produced by exposure to radiation or test compounds can be detected as an inhibition of cell outgrowth. - [Read Human Oesophageal Culture Protocol]

Human Oesophageal Culture Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/28.htm#TOP

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

Protocol describes a method for establishing short-term explant cultures of oesophageal mucosa. Adverse effects produced by exposure to radiation or test compounds can be detected as an inhibition of cell outgrowth. - [Read Human Oesophageal Culture Protocol]

Human Skin Fibroblast/Collagen Lattice Cytotoxicity Test - http://embryo.ib.amwaw.edu.pl/invittox/prot/27.htm#TOP

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

Skin fibroblasts are incorporated into 3-D collagen lattices containing the test compounds. An inhibition of lattice contraction indicates a possible toxic effect which is verified by trypan blue exclusion for cell viability. - [Read Human Skin Fibroblast/Collagen Lattice Cytotoxicity Test]

Human Thyroid Culture Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/29.htm#TOP

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

This method enables the culturing of thyroid cells without loss of differentiation and medium change. It is potentially useful for the long-term study of drug effects on the thyroid gland. - [Read Human Thyroid Culture Protocol]

Human Thyroid Culture Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/29.htm#TOP

Category: Pharmacology and Toxicology Protocols: Drug Discovery

Protocol enables the culturing of thyroid cells without loss of differentiation and medium change. It is potentially useful for the long-term study of drug effects on the thyroid gland. - [Read Human Thyroid Culture Protocol]

In Vitro Micromass Teratogen Assay - http://embryo.ib.amwaw.edu.pl/invittox/prot/114.htm

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

The effect of a test compound, in the presence and absence of S-9 mix, on the differentiation and growth of rat limb bud and CNS cells in vitro indicates whether it is potentially a teratogen in vivo. - [Read In Vitro Micromass Teratogen Assay]

In Vitro Model For Studies Of Prostagland H Synthase (PHS)- Mediated Genotoxicity Of Xenobiotics - http://embryo.ib.amwaw.edu.pl/invittox/prot/61.htm

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

This protocol describes the use of SEMV cells (a cell line derived from ram seminal vesicles) in studies into prostaglandin H synthase-mediated metabolism of xenobiotics in intact cells. - [Read In Vitro Model For Studies Of Prostagland H Synthase (PHS)- Mediated Genotoxicity Of Xenobiotics]

In Vitro Prediction of the Maximum Tolerated Dose Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/66.htm

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

The results of cytotoxicity tests in primary cultures of rat hepatocytes and in MDBK and McCoy cells can be used to predict the in vivo 4-wk maximum tolerated dose in rats and dogs. A correlation between in vitro cytotoxicity, as measured in this system, and LD50 values in rats and mice has also been established. - [Read In Vitro Prediction of the Maximum Tolerated Dose Protocol]

Injection of dsRNA and Electroporation in Avian Embryos Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4504

Category: Avian Bird Protocols

Injection of dsRNA and Electroporation in Avian Embryos Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4504

Category: RNAi Technology Protocols: RNAi Avian Embryos Protocols

Protocol describes a method for in ovo transfection of avian embryos with double-stranded RNA (dsRNA). The dsRNA is injected into the spinal cord of the embryo. Subsequent electroporation facilitates the cellular uptake of the dsRNA molecules. It may be necessary to optimize the stage of the embryo and the electroporation procedure to improve the effectiveness of in ovo RNAi—cell competence changes with differentiation. - [Read Injection of dsRNA and Electroporation in Avian Embryos Protocol]

Invitro Model for Prostoglandin H Synthase (PHS) Mediated Genotoxicity of Xenobiotics - http://embryo.ib.amwaw.edu.pl/invittox/prot/61.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

Protocol describes the use of SEMV cells (a cell line derived from ram seminal vesicles) in studies into prostaglandin H synthase-mediated metabolism of xenobiotics in intact cells. - [Read Invitro Model for Prostoglandin H Synthase (PHS) Mediated Genotoxicity of Xenobiotics]

Isolated Rat Glomeruli and Proximal Tubules - http://embryo.ib.amwaw.edu.pl/invittox/prot/5.htm

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

The basis of this procedure is that two specific cell type preparations may be isolated, exposed separately to various compounds over a range of concentrations, and the cytotoxicity of these determined. Parameters deemed indicative of a cytotoxic effect include a reduction in de novo protein synthesis and decreased glucose and fatty acid metabolism. A cytotoxic effect may indicate that a chemical is likely to be nephrotoxic in vivo. - [Read Isolated Rat Glomeruli and Proximal Tubules]

Isolation of Rat Hepatocytes Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/20.htm

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

Collagenase perfusion of rat liver yields a hepatocyte suspension which may be exposed to test compounds in order to assess their effects on cell viability and enzyme leakage. - [Read Isolation of Rat Hepatocytes Protocol]

Liver Slice Hepatotoxicity Screening System Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/42.htm

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

Leakage of lactate dehydrogenase and alanine aminotransferase from rat and mouse liver slices exposed to the test compound is used as a measure of hepatotoxicity. - [Read Liver Slice Hepatotoxicity Screening System Protocol]

LS-L929 Cytotoxcitiy Test - http://embryo.ib.amwaw.edu.pl/invittox/prot/38.htm

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

This simple cell culture-based cytotoxicity test (in which cell viability is determined by uptake of the dyes ethidium bromide and fluorescein acetate) has been developed as a general test for acute toxicity. - [Read LS-L929 Cytotoxcitiy Test]

Lung Cell Assay Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/48.htm

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

Potential embryotoxicity is assessed by monitoring the effect of the test compound on total protein synthesis, and DNA synthesis in cultured human foetal lung fibroblasts. Rat lung epithelial cells can be used to determine cytotoxicity of select compounds because of their ability to metabolise xenobiotics. - [Read Lung Cell Assay Protocol]

Membrane Permability as a Measure of Cytotoxcity in Perfused Cell Cultures - http://embryo.ib.amwaw.edu.pl/invittox/prot/9.htm

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

Membrane permeability of perfused cell cultures, as determined by the efflux of [3H]-2-deoxy-D-glucose-6phosphate, is used as an indicator of the cytotoxic effect of chemicals. - [Read Membrane Permability as a Measure of Cytotoxcity in Perfused Cell Cultures]

Mouse Caesarean Section and Fostering Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4381

Category: Mouse Protocols: Mouse Reproductive System Surgery Protocols

Protocol describes a method for Caesarean section and fostering. Caesarean section is required if the recipient of an embryo transfer or any pregnant mouse has not given birth by the delivery time normal for the particular strain. - [Read Mouse Caesarean Section and Fostering Protocol]

Mouse Embryo Transfer into Pseudopregnant Recipient Protocol - http://members.cox.net/microinjectionworkshop/technical/tnp2.html

Category: Mouse Protocols: Mouse Embryology Protocols

1-cell embryo transfer into pseudopregnant recipient female mouse. - [Read Mouse Embryo Transfer into Pseudopregnant Recipient Protocol]

MOUSE EMBRYONIC FIBROBLASTS PDF - http://www.nki.nl/nkidep/h8/Groups/teriele/protocols/2.pdf

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: MEF Protocols Mouse Embryonic Fibroblasts

Preparation of primary mouse embryonic fibroblasts, Preparation of mitotically inactive mouse embryo fibroblasts, Irradiation of the MEFs. Nederlands Kanker Instituut Teriele. - [Read MOUSE EMBRYONIC FIBROBLASTS PDF]

Native Aequorin - http://www.luxbiotech.com/Docs/Nanolight/Aequorin.pdf

Category: Signalling Signal Transduction Protocols: Calcium Measurement Protocols

Native Aequorin. NanoLight Technology. Aequorin has advantages over other Ca2+ indicators, for example, low leakage rate from cells, lack of intracellular compartmentalization or sequestration and it does not disrupt cell functions or embryo development. - [Read Native Aequorin]

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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