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Derivation of Mouse Embryonic Fibroblasts (MEF) Part I PDF - http://www.wicell.org/uploads/media/Protocols_Part_1_05.pdf

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

WiCell Research Institute. Recipes, Frequently Asked Questions, Mouse Embryo Extraction and Tissue, Harvesting Mouse Embryonic Fibroblasts (MEF)from mice. - [Read Derivation of Mouse Embryonic Fibroblasts (MEF) Part I PDF]

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]

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]

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]

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]

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]

Autoradiography for mRNA detection in. mouse embryo tissue sections. PDF - http://www.oup.co.uk/pdf/pas/12-3-9.pdf

Category: Radioactivity: Autoradiography

Antonio Simeone. MRC Centre for Developmental Neurobiology, New Huntâ€™s Houre. - [Read Autoradiography for mRNA detection in. mouse embryo tissue sections. PDF]

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]

Blastocyst Embryo Transfer into Pseudopregnant Mouse Protocol - http://members.cox.net/microinjectionworkshop/technical/tnp6.html

Category: Mouse Protocols: Mouse Embryology Protocols

Protocol for blastocyst embryo transfer into pseudopregnant recipient female mouse. - [Read Blastocyst Embryo Transfer into Pseudopregnant Mouse Protocol]

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]

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]

Agarose Overlay Assay Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/31.htm

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

Protocol descibes the use of L929 mouse fibroblast cells cultured in vitro in an agarose overlay assay to assess the toxicity of test substances. The assay may be useful in assessing the irritation potential of test substances (e.g. surfactant-based products) as an alternative to the Draize rabbit eye test. - [Read Agarose Overlay Assay 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]

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]

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]

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]

Aggregation of ES Cells with Mouse Morula- Stage Embryos Protocol - http://fds.oup.com/www.oup.co.uk/pdf/pas/13-10-6.pdf

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Protocol for the aggregation of ES cells with mouse morula-stage embryos. Includes: Preparation of aggregation walls; ES cell preparation; Embryo preparation and aggregation. - [Read Aggregation of ES Cells with Mouse Morula- Stage Embryos Protocol]

Drosophila Embryo Microinjection Protocol - http://vosshall.rockefeller.edu/protocols/DrosophilaEmbryo.pdf

Category: Drosophila Protocols

Protocol for Drosophila embryo microinjection. - [Read Drosophila Embryo Microinjection Protocol]

FISH Protocols for Drosophila - http://www.utoronto.ca/krause/FISH.html

Category: Drosophila Protocols: Drosophila Genetics Protocols

FISH protocols for Drosophila. Includes: RNA Probe Preparation; Embryo Collection and Fixation; Single FISH on Drosophila embryos; Post-Fixation, Hybridization and Post-Hybridization Washes; Development of FISH Signal; Storage, Mounting and Viewing of Samples; Double FISH on Drosophila Embryos; RNA-Protein Double Labeling; FISH on Dissected Tissues. - [Read FISH Protocols for Drosophila]

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]

Embryo Lysates & Immunoprecipitation Protocol - http://spot.colorado.edu/~klym/Methods/Precipitation.htm

Category: Xenopus Protocols: Xenopus Embryo Manipulation Protocols

Protocol for embryo lysates and immunoprecipitation: protein A agarose & protein A magnetic beads. Includes: Embryo lysates; Immunoprecipitation with protein A agarose; Immunoprecipitation with protein A magnetic beads for silver stain/sequencing analysis. - [Read Embryo Lysates & Immunoprecipitation Protocol]

Clearing Arabidopsis Ovule Materials with the Hoyer's Solution Protocol - http://www.hybtech.org/Liu/Hoyer_clearing.htm

Category: Plant Biology Protocols

Protocol can be used for clearing intact seeds of arabidopsis, which can then be observed under Normarski optics. This is an efficient way to analyse embryo and endosperm development without sectioning. - [Read Clearing Arabidopsis Ovule Materials with the Hoyer's Solution 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]

Cell Culture Phototoxicity Test Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/44.htm

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

Human A431 cells and mouse 3T3 cells are exposed in culture to UV light both in the presence and absence of test compound. Phototoxicity is expressed as a decrease in cell viability as determined by the MTT assay. - [Read Cell Culture Phototoxicity Test Protocol]

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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