User Panel

My Panel

Bookmark Science Articles

line Protocols

Search results for: line

Protocols » Search Results

Search Results Protocol Links Sort by: Hits | Alphabetical

A Targeted Screen to Detect Recessive Mutations that have Quantitative Effects Protocol - http://www.nervenet.org/papers/Consomic.html

Category: Genetics and Genomics: Genotyping Protocols: Mutation Detection Protocols

Describes an experimental cross in mice that can be used to define and map induced germ-line mutations that map to a single chromosome. The cross is a modification and extension of a conventional three-generation recessive mutagenesis screen. Includes: The Mutagenesis Breeding Plan; Consomic Strains; Generating Mutations; Generating and Genotyping G2 Females; Genotyping G3 Progeny; Phenotyping G4 Progeny; etc.. - [Read A Targeted Screen to Detect Recessive Mutations that have Quantitative Effects Protocol]

Adapting Cells to a Serum-Free Environment Protocol - https://fscimage.fishersci.com/webimages_FSC/downloads/HyClone_Protocol_8.pdf

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

There are many ways to adapt cell lines to serum-free media. Five methods are presented that are designed for adapting hybridomas to a protein-free medium. These protocols may require some modifications for your particular cell line and conditions. - [Read Adapting Cells to a Serum-Free Environment Protocol]

Antisense Oligonucleotide Transfection of RAW 264.7 Cells with FuGENE 6 in a 24-Well Dish - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000261.pdf?pid=PP00000261

Category: Transfection Protocols: Stable Transfection Protocols

The AfCS is utilizing antisense technology to manipulate signaling protein expression in the RAW 264.7 macrophage-like cell line. This can be achieved by the transfection of gene-specific antisense oligonucleotides (ASOs). The following procedure involves the transfection of ASOs into RAW 264.7 cells using FuGENE 6 transfection reagent. Subsequently, the isolated total RNA or protein from these transfected cells can be used to assess the level of mRNA or protein knockdown, respectively. - [Read Antisense Oligonucleotide Transfection of RAW 264.7 Cells with FuGENE 6 in a 24-Well Dish]

B- Galactosidase Assay Protocol - http://www.biochem.northwestern.edu/ibis/morimoto/research/Protocols/VI.%20Transcription/I.%20b-gal%20.pdf

Category: Reporter Gene Assays: B-Galactosidase Assay Protocols

The recommended amount of RSV-ß-Galactosidase plasmid to use for transfection of cells (60 mm or 100 mm dish) is 1-2 µg. The optimal amount of plasmid DNA will be determined by the efficiency of transfection , which is very dependent upon the particular cell line and transfection protocol. - [Read B- Galactosidase Assay Protocol]

Calcium Phosphate Transfection Method - http://www.flemingtonlab.com/Protocols/CalciumPhosphateTransf.pdf

Category: Transfection Protocols: Calcium Phosphate Transfection Protocols

This calcium phosphate transfection method works best in cell lines that are 1) highly transformed and 2) adherent (Hela, U2OS, SAOS2, AdAH, NPC-KT and obtain from 20% to 100% transfection efficiency depending on the cell line). Works well for transient experiments but precautions should be used in the design and interpretation of experiments based on the discussion below. Also works very well for generating stable cell lines. This method is quite sensitive to the amount of input plasmid. - [Read Calcium Phosphate Transfection Method]

Calcium-phosphate-mediated Transfection of Eukaryotic Cells with Plasmid DNAs - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3871

Category: Transfection Protocols: Calcium Phosphate Transfection Protocols

Calcium phosphate forms an insoluble precipitate with DNA, which attaches to the cell surface and is taken into the cells by endocytosis. The protocol is easily adapted for use with other types of cells, both adherent and nonadherent. This protocol is a modified version of a method published by Jordan et al. (1996) who rigorously optimized calcium-phosphate-based transfection methods for Chinese hamster ovary cells and the 293 line of human embryonic kidney cells. - [Read Calcium-phosphate-mediated Transfection of Eukaryotic Cells with Plasmid DNAs]

Cell Lysate Extracts - http://www.upstate.com/misc/protocol_detail.q.prot.e.CellLysate_Prep.a.name.e.Cell_Lysate_Extracts-General_Protocols

Category: Protein Protocols: Protein Extraction From Cells

Cell Lysate Extracts. Great protocols for cell lysis preparation from a variety of cell types. There are numerous methods of cell stimulation and lysis. For a given protein, Upstateâ€™s Laboratories determine the specific treatment upon initial testing of its products. It is important to select the correct cell line, stimulation procedure (if any), and lysis protocol. Upstate. - [Read Cell Lysate Extracts]

Cell Thawing and cell line Freezing Protocol - http://iacf.bsd.uchicago.edu/Monoclonal_Antibody_Facility/FreezeThawProtocol.htm

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

Method for freezing and thawing cells. Immunology Applications Core Facility. U. Chicago - [Read Cell Thawing and cell line Freezing Protocol]

Culturing Trophoblast Stem (TS) Cell Lines Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4406

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Culture conditions have been established for a second blastocyst-derived cell line, trophoblast stem (TS) cells, in addition to embryonic stem (ES) cells. This protocol describes a method for culturing TS cell lines. These cells can then be used to study trophoblast differentiation and placental function. - [Read Culturing Trophoblast Stem (TS) Cell Lines Protocol]

CYP1A1-Inducing Potency and Cytotoxicity Test in the HEPA-1 Mouse Hepatoma Cell Line - http://embryo.ib.amwaw.edu.pl/invittox/prot/112.htm

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

This bioassay utilizes cultured Hepa-lclc7 (Hepa-1) mouse hepatoma cells to assess the CYPlA1-inducing potency or cytotoxicity of pure test chemicals or environmental samples. In the Hepa-l induction test , the CYPlA1-inducing potency of the test sample is detected as increased aryl hydrocarbon hydroxylase (AHH) and 7-ethoxyresorufin O-deethylase (EROD) activities. - [Read CYP1A1-Inducing Potency and Cytotoxicity Test in the HEPA-1 Mouse Hepatoma Cell Line]

Cytokine Bioassays Protocols - http://www.ebioscience.com/ebioscience/appls/BAC.htm?

Category: Signalling Signal Transduction Protocols: Cytokine Protocols

Cytokine-Induced Proliferation of Indicator Cell Lines, TNF-Î±-Induced Killing of L929 Cell Line, IFN-Î³ Protection from Viral Infection of Cell Lines. eBiosciences. - [Read Cytokine Bioassays Protocols]

Cytokine Neutralization, In Vitro Using Antibodies - http://www.ebioscience.com/ebioscience/appls/NU.htm

Category: Signalling Signal Transduction Protocols: Cytokine Protocols

Antibody Neutralization of TNF-Î±-Induced Killing of L929 Cell Line, IFN-Î³-Protection from Viral Infection of L929 and A549 Cell Lines, TNF-Î±-Induced Killing of L929 Cell Line. eBiosciences. - [Read Cytokine Neutralization, In Vitro Using Antibodies]

Dose of G418 to be used for selection - http://invitrogen.custhelp.com/cgi-bin/invitrogen.cfg/php/enduser/std_adp.php?p_sid=rLTNWTng&p_lva=&p_faqid=80

Category: Cell Biology and Cell Culture: Cell Transfection Protocol

What dose of Geneticin (G418) should I use for stable cell line selection? Invitrogen FAQs. - [Read Dose of G418 to be used for selection]

Generation of Growth Curve Protocol - https://fscimage.fishersci.com/webimages_FSC/downloads/HyClone_Protocol_4.pdf

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

Generation of a growth curve can be useful in evaluating the growth characteristics of a cell line. From a growth curve, the lag time, population doubling time, and saturation density can be determined. - [Read Generation of Growth Curve Protocol]

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]

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]

Isolation of Retroelement from Plant Genomic DNA - http://www.le.ac.uk/biology/phh4/retros.htm

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

Retroelements and their derivatives are a ubiquitous and abundant component of plant genomes. Major classes of retroelements include the Pseudoviridae (Ty1-copia ), the Metaviridae (Ty3 -gypsy) and the Retroposineae LINE (non-LTR) groups. All reverse transcribing elements can be included in a universal classification. Includes: Pseudoviridae (Ty1-copia) Degenerate Primers; Metaviridae (Ty3-gypsy) Element Degenerate Primers; LINE Element Degenerate Primers; PCR Programmes. - [Read Isolation of Retroelement from Plant Genomic DNA]

Karyotyping Protocol - http://humgen.wustl.edu/hdk_lab_manual/cam/cam5.html

Category: Cytogenetics Protocols

Karyotyping is a valuable research tool used to determine the chromosome complement within cultured cells. It is important to keep in mind that karyotypes evolve with continued culture. Because of this evolution, it is important for the interpretation of biochemical or other data, that the karyotype of a specific sub-line be determined. - [Read Karyotyping Protocol]

Optimization Protocol â€“ Cell Line Electroporation PDF - http://www.eppendorf.com/script/cms-newspic.php?id=4298&inline=1&col=DOWNLOADFILE

Category: Cell Biology and Cell Culture: Cell Electroporation Protocols

Optimization of Electroporation Transfection of DNA Protocol into Cell Lines. Eppendorf - [Read Optimization Protocol â€“ Cell Line Electroporation PDF]

Passage Procedure for RAW 264.7 Cells - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000159.pdf?pid=PP00000159

Category: Cell Culture Protocols: Cell Lines Protocols

RAW 264.7 cells are a macrophage-like, Abelson leukemia virus transformed cell line derived from BALB/c mice. For routine maintenance in culture (passage), cells are seeded at a confluence of approximately 10% (1 x 106 and 3 x 106 cells in 100-mm and 150-mm plates, respectively) and grown to a confluence of approximately 80%. This procedure requires the cells to be split every two days. - [Read Passage Procedure for RAW 264.7 Cells]

Preparation of Endothelial Cells Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=E538DDD4A302A0DF2CDDD9363072C1CA&objectid=6673B460F50B90057960CE47ABB708A4

Category: Cell Biology and Cell Culture: Endothelial Cell Protocols

Endothelial cells, which line blood vessels, can be prepared from a variety of tissues. They are frequently prepared from the umbilical vein, which is relatively easy to obtain. The procedure is clearly described and provides a large population of highly purified endothelial cells. There are also methods for obtaining endothelial cells from other tissues such as fat, skin, and mucosa. These methods require special care and generate smaller populations of cells. - [Read Preparation of Endothelial Cells 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]

Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4484

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the culture of embryonic stem (ES) cells using mitotically inactivating primary mouse embryonic fibroblast (MEF) cells as a feeder layer (preparation described in Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates). The ES culture medium is supplemented with recombinant leukemia inhibitory factor (LIF) to help maintain the cells as pluripotent stem cells. This protocol has been optimized for the ES-D3 cell line. - [Read Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol]

Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4484

Category: Stem Cell Protocols: Mouse Embryonic Fibroblasts Protocols

Articles (1-10 of 16)

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.

Genomic DNA Labeling of Microarrays Protocol

DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.

Drosophila Cell Transfection Protocol

A simple Drosophila tissue culture cell transfection method.

Tubulin Polymerization Protocol using GTP and GMPCPP

Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.

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.

Acid Washing Microinjection Pipettes Protocol

Acid Washing of Glass microinjection pipettes protocol.

Paraffin Embedding Protocol

Paraffin Embedding Protocol for molecular profiling. This Paraffin Embedding Protocol describes the processing of the tissues into sections following ethanol fixation. Molecular profiling (MP) is a technique that is used to visualize the global patterns of RNA expression or protein expression in various cell types and disease processes.

In Vitro Translated Xenopus Mos Kinase Assay Protocol

In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.