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Antibiotic and Selection Reagent Usage for Resistant E. coli Cells PDF - http://www.atcc.org/pdf/AntibioticTable.pdf

Category: Microbiology: Antibiotics Concentration in Media LB

Antibiotics, ampicillin, choramphenicol, G418, X-Gal and IPTG also stock concentrations. ATCC. - [Read Antibiotic and Selection Reagent Usage for Resistant E. coli Cells PDF]

Antibody Addition and Detection for Staining Caenorhabditis elegans Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4523

Category: C. elegans Protocols: C. elegans Staining Protocols

Extreme care should be used to identify and verify positive reactions, however, because cross-reactions are common. Counterstaining is essential for examining worms by immunofluorescence and is used to identify the exact cell in which an antigen appears. Methods for counterstaining include labeling all cells with a fluorescent dye that is specific for nucleic acids (e.g., DAPI or propidium iodide) and using GFP driven by tissue-specific promoters. - [Read Antibody Addition and Detection for Staining Caenorhabditis elegans 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]

Apoptosis Detection: TUNEL Protocol - http://biochem.dental.upenn.edu/~sdmfacs/apopproc.htm#brduapop

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

Protocol for apoptosis detection using TUNEL protocol. This technique uses the enzyme terminal deoxynucleotidyl transferase (TdT) to label cells that have oligonucleosomal nicks/strand breaks in their DNA. - [Read Apoptosis Detection: TUNEL Protocol]

Application of Direct Current Electric Fields to Cells and Tissues in vitro - http://www.natureprotocols.com/2006/12/12/application_of_direct_current.php

Category: Cell Biology and Cell Culture

Protocol applies EFs to cells in vitro but has been modified and to use electrotactic chambers to accommodate cells growing in planar culture or in three-dimensional (3D) gels, en bloc tissue cultures in 3D and possible small embryos, such as that from frog and zebra fish. The EF is applied to the cells or tissues cultured in a customer designed electrotactic chamber via agar salt bridges, Steinbergâ€™s solution and Ag/AgCl electrodes. - [Read Application of Direct Current Electric Fields to Cells and Tissues in vitro]

AS Ammonium Sulfate Binding Assay for Cells - http://www.hopkinsmedicine.org/cellbio/devreotes/as.htm

Category: Immunology: Chemotaxis

AS Ammonium Sulfate Binding Assay for Cells. Devreotes Lab, John Hopkins Medical Centre. - [Read AS Ammonium Sulfate Binding Assay for Cells]

Assay for ß-galactosidase in Extracts of Mammalian Cells - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3952

Category: Reporter Gene Assays: B-Galactosidase Assay Protocols

The assay for ß-galactosidase relies on the ability of the enzyme to catalyze the hydrolysis of ONPG (o-nitrophenyl-ß-D- galactopyranoside) to free o-nitrophenol, which absorbs light at 420 nm. In this protocol, extracts of cells transfected with a ß-galactosidase reporter plasmid are incubated with ONPG. - [Read Assay for ß-galactosidase in Extracts of Mammalian Cells]

Assay for ß-galactosidase in Extracts of Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3952

Category: Yeast Protocols: Yeast Genetics Protocols: B-Galactosidase Assay Protocols

In this protocol, extracts of cells transfected with a ß-galactosidase reporter plasmid are incubated with ONPG. - [Read Assay for ß-galactosidase in Extracts of Mammalian Cells Protocol]

Assay for Luciferase in Extracts of Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3642

Category: Reporter Gene Assays: Luciferase Assay Protocols

In this protocol, cells transfected with a luciferase reporter plasmid are lysed in a detergent-containing buffer. Luciferase in the extract catalyzes an oxidation reaction in which D-luciferin is converted to oxyluciferin, with production of light at 556 nm that can be quantified in a luminometer. - [Read Assay for Luciferase in Extracts of Mammalian Cells Protocol]

Assay of ß-Galactosidase in Yeast: Assay of Crude Extracts - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4157

Category: Reporter Gene Assays: B-Galactosidase Assay Protocols

Method describes how a crude extract is prepared, and the activity is normalized to the amount of protein assayed. This method is particularly suitable for comparing cells that are grown under very different conditions or that have different genetic backgrounds. - [Read Assay of ß-Galactosidase in Yeast: Assay of Crude Extracts]

Assay of ß-Galactosidase in Yeast: Assay of Crude Extracts Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4157

Category: Yeast Protocols: Yeast Genetics Protocols: B-Galactosidase Assay Protocols

Protocol describes how a crude extract is prepared, and the activity is normalized to the amount of protein assayed. This method is particularly suitable for comparing cells that are grown under very different conditions or that have different genetic backgrounds. - [Read Assay of ß-Galactosidase in Yeast: Assay of Crude Extracts Protocol]

Assay of ß-Galactosidase in Yeast: Freeze/Thaw Assay by Chemiluminescence in 96-Well Plates - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4160

Category: Reporter Gene Assays: B-Galactosidase Assay Protocols

Method described indicates how cells are lysed by a simple freeze/thaw protocol and treated with a chemiluminescent substrate. - [Read Assay of ß-Galactosidase in Yeast: Freeze/Thaw Assay by Chemiluminescence in 96-Well Plates]

Assay of ß-Galactosidase in Yeast: Freeze/Thaw Assay by Chemiluminescence in 96-Well Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4160

Category: Yeast Protocols: Yeast Genetics Protocols: B-Galactosidase Assay Protocols

In this protocol the cells are lysed by a simple freeze/thaw protocol and treated with a chemiluminescent substrate. - [Read Assay of ß-Galactosidase in Yeast: Freeze/Thaw Assay by Chemiluminescence in 96-Well Protocol]

Assay of ß-Galactosidase in Yeast: Freeze/Thaw Assay by Chemiluminescence in Microcentrifuge Tubes - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4159

Category: Reporter Gene Assays: B-Galactosidase Assay Protocols

In the method described here, the cells are lysed by a simple freeze/thaw protocol and . . . - [Read Assay of ß-Galactosidase in Yeast: Freeze/Thaw Assay by Chemiluminescence in Microcentrifuge Tubes]

Assay of ß-Galactosidase in Yeast: Freeze/Thaw Assay by Chemiluminescence in Microcentrifuge Tubes - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4159

Category: Yeast Protocols: Yeast Genetics Protocols: B-Galactosidase Assay Protocols

In this protocol the cells are lysed by a simple freeze/thaw protocol. - [Read Assay of ß-Galactosidase in Yeast: Freeze/Thaw Assay by Chemiluminescence in Microcentrifuge Tubes]

Assay of ß-Galactosidase in Yeast: Permeabilized Cell Assay - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4158

Category: Reporter Gene Assays: B-Galactosidase Assay Protocols

In the method described here, the cells are permeabilized to allow the substrate to enter the cells and the activity is normalized to the number of cells assayed. - [Read Assay of ß-Galactosidase in Yeast: Permeabilized Cell Assay]

Assay of ß-Galactosidase in Yeast: Permeabilized Cell Assay Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4158

Category: Yeast Protocols: Yeast Genetics Protocols: B-Galactosidase Assay Protocols

In this protocol, the cells are permeabilized to allow the substrate to enter the cells and the activity is normalized to the number of cells assayed. - [Read Assay of ß-Galactosidase in Yeast: Permeabilized Cell Assay Protocol]

Assay of Cyclic AMP in Lysates of Cells - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000015.pdf?pid=PP00000015

Category: Signalling Signal Transduction Protocols

Protocol allows you to measure the content of cyclic adenosine 3',5'-monophosphate (cyclic AMP or cAMP) in splenic B lymphocytes (B cells) in an enzyme-linked immunoassay. This protocol utilizes acetylation of cAMP to improve sensitivity and reduce interference. Protocol includes information on: how to determine cAMP, calculations and reagents and materials. - [Read Assay of Cyclic AMP in Lysates of Cells]

Assay of Intracellular Free Calcium in RAW 264.7 Cells - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000210.pdf?pid=PP00000210

Category: Signalling Signal Transduction Protocols: Calcium Measurement Protocols

This protocol describes a method to assess concentrations of free cytoplasmic calcium, [Ca2+]i, in cultured adherent RAW 264.7 cells, using a 96-well plate format. This objective is accomplished by using the Ca2+-sensitive fluorescent dye, fluo-3, which permeates cell membranes as an ester and is hydrolyzed in the cell to its Ca2+-sensitive acidic form. - [Read Assay of Intracellular Free Calcium in RAW 264.7 Cells]

Assay of Intracellular Free Calcium in RAW 264.7 Cells for Ligand Screen Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000176.pdf?pid=PP00000176

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Intracellular Ion Measurement Protocols

This protocol describes a method to assess concentrations of free cytoplasmic calcium (Ca2+) in cultured RAW 264.7 cells. This objective is accomplished with the Ca2+-sensitive fluorescent dye, fluo-3, which permeates cells as an ester and is hydrolyzed in the cell to its Ca2+-sensitive acidic form. Fluorescence is measured over time with adherent cells that have been washed free of extracellular dye. - [Read Assay of Intracellular Free Calcium in RAW 264.7 Cells for Ligand Screen Protocol]

Assay of Intracellular Free Calcium in RAW 264.7 Cells Loaded with Fluo-3 Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000210.pdf?pid=PP00000210

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Intracellular Ion Measurement Protocols

This protocol describes a method to assess concentrations of free cytoplasmic calcium, [Ca2+]i, in cultured adherent RAW 264.7 cells, using a 96- well plate format. This objective is accomplished by using the Ca2+-sensitive fluorescent dye, fluo-3, which permeates cell membranes as an ester and is hydrolyzed in the cell to its Ca2+-sensitive acidic form. Fluorescence for the adherent cells is measured over time by using a bottom read of a 96-well plate, with cells that have been washed. - [Read Assay of Intracellular Free Calcium in RAW 264.7 Cells Loaded with Fluo-3 Protocol]

Assay of Intracellular Free Calcium in RAW 264.7 Cells Loaded with Fura-2 (with FLEXstation) - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000211.pdf?pid=PP00000211

Category: Signalling Signal Transduction Protocols: Calcium Measurement Protocols

This protocol describes a method to assess concentrations of free cytoplasmic calcium, [Ca2+], in cultured adherent RAW 264.7 cells, using a 96-well plate format. This objective is accomplished by using the Ca2+-sensitive fluorescent dye, fura-2, which permeates cell membranes as an ester and is hydrolyzed in the cell to its Ca2+-sensitive acidic form. - [Read Assay of Intracellular Free Calcium in RAW 264.7 Cells Loaded with Fura-2 (with FLEXstation)]

Assay of Intracellular Free Calcium in RAW 264.7 Cells Loaded with Fura-2 Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000211.pdf?pid=PP00000211

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Intracellular Ion Measurement Protocols

This protocol describes a method to assess concentrations of free cytoplasmic calcium, [Ca2+]i , in cultured adherent RAW 264.7 cells, using a 96- well plate format. This objective is accomplished by using the Ca2+-sensitive fluorescent dye, fura-2, which permeates cell membranes as an ester and is hydrolyzed in the cell to its Ca2+-sensitive acidic form. - [Read Assay of Intracellular Free Calcium in RAW 264.7 Cells Loaded with Fura-2 Protocol]

Assay of Intracellular Free Calcium in Suspended B Cells - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000011.pdf?pid=PP00000011

Category: Signalling Signal Transduction Protocols: Calcium Measurement Protocols

This protocol describes a method to assess concentrations of free cytoplasmic calcium (Ca2+) in mouse splenic B cells in the absence and presence of ligands for cell surface receptors. This objective is accomplished with the Ca2+-sensitive fluorescent dye, Fluo-3, which permeates cells as an ester and is hydrolyzed in the cell to its Ca2+ sensitive acidic form. - [Read Assay of Intracellular Free Calcium in Suspended B Cells]

Assay of Intracellular Free Calcium in Suspended B Cells Dual Ligand PDF - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000140.pdf?pid=PP00000140

Category: Signalling Signal Transduction Protocols: Calcium Measurement Protocols

The following protocol is a method to measure concentrations of free cytoplasmic calcium (Ca2+) in mouse splenic B cells in the presence of two ligands. Signalling Gateway - [Read Assay of Intracellular Free Calcium in Suspended B Cells Dual Ligand PDF]

Articles (1-10 of 24)

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.

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.

Microinjection Pipettes Protocol

Microinjection Pipettes Preparation Protocol.

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.

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