induced Protocols

Category: Cell Biology and Cell Culture: Apoptosis Protocols

A Photoelectric Method for analyzing NO-induced apoptsis. Zhang et al. 2000 - [Read A Photoelectric Method for analyzing NO-induced apoptsis.]

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]

Category: Mouse Protocols: Assembling Chimeras Mouse Protocols

Protocol describes a method for producing diploid embryo-tetraploid embryo chimeras. It requires the timed combination of four-cell-stage tetraploid embryo production and the procedure for diploid embryo-diploid embryo aggregation. The resulting chimeras are useful for phenotypic analysis when an induced mutation has an extraembryonic phenotype. - [Read Assembling Aggregates between Diploid and Tetraploid Embryos Protocol]

Category: Neuroscience Protocols

Bead aggregation assay to demonstrate presynaptic differentiation induced by the NGL family of cell adhesion molecules. - [Read Bead Aggregation Assay Protocol]

Category: Signalling Signal Transduction Protocols: Cytokine Protocols

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

Category: Signalling Signal Transduction Protocols: Cytokine Protocols

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

Category: Plant Biology Protocols: Plant Genetics Protocols

Virus-induced gene silencing (VIGS) uses a virus to deliver a sequence from a gene of interest into a host plant. The virus carrying the fragment of the gene of interest must be capable of replication if dsRNA is to be produced. One or two leaves are inoculated with Agrobacterium strains carrying the VIGS vector possessing the gene fragment. The virus then replicates and spreads throughout the plant, mediating silencing. - [Read Delivery of dsRNA into Plants by VIGS Methodology]

Category: RNAi Technology Protocols: RNAi Mammalian Cells Protocols

Protocol describes the use of western blotting to analyze the RNAi-induced depletion of target protein from mammalian cells.  Immunofluorescence can be used as an alternative. - [Read Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting Protocol]

Category: Western Blot Protocols

Protocol describes the use of western blotting to analyze the RNAi-induced depletion of target protein from mammalian cells.  Immunofluorescence can be used as an alternative. - [Read Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting Protocol]

Category: Yeast Protocols: Yeast Protein Protocols

This assay is performed to detect ubiquitylated proteins in yeast. Yeast that have been transformed with a vector expressing polyhistidine-tagged ubiquitin (Ub) under the control of a copper-inducible promoter are grown, induced with copper, and harvested. Total ubiquitylated proteins are then recovered by nickel-affinity chromatography, and specific proteins are detected by Western blotting. - [Read Detection of Ubiquitylated Proteins in Yeast Protocol]

Category: Immunology: Chemotaxis

GTPγS-induced Inhibition of Binding. Chemotaxis assay protocols. Devreotes Lab, John Hopkins Medical Centre. - [Read GTPγS-induced Inhibition of Binding]

Category: Signalling Signal Transduction Protocols

This protocol assays inhibition of in vivo binding of [3H]-cAMP to cAR1 by GTPγS. Dictyostelium discoideum respond to extracellular cAMP through the cAMP chemoattractant receptor (cAR1). Binding of cAMP to the G protein-coupled cAR1 is inhibited by the GTP analog GTPγS. Protocol includes information on: Solutions used, BioReagents and Chemicals and Protocol Hints. - [Read GTPγS-Induced Inhibition of cAMP Binding to the cAMP Receptor (cAR1) in Dictyostelium Discoideum]

Category: Immunology: Immunofluorescence Protocols

This protocol describes the use of a specific antibody that recognizes the targeted gene product to detect RNAi-induced gene knockdown in mammalian cells. Western blot technology can be used as an alternative (see Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting). - [Read Immunofluorescence Detection of RNAi-Induced Protein Knockdown in Mammalian Cells Protocol]

Category: RNAi Technology Protocols: RNAi Mammalian Cells Protocols

Protocol describes the use of a specific antibody that recognizes the targeted gene product to detect RNAi-induced gene knockdown in mammalian cells.  Western blot technology can be used as an alternative. - [Read Immunofluorescence Detection of RNAi-Induced Protein Knockdown in Mammalian Cells Protocol]

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

In this protocol, a bacterial lysogen is constructed from a recombinant bacteriophage {lambda} encoding a fusion protein of interest. The resulting lysogenic colonies are induced to synthesize the fusion protein, which is then isolated in preparation for functional and biochemical analyses. - [Read Preparation of Lysates Containing Fusion Proteins Encoded by Bacteriophage {lambda} Lysogens]

Category: Immunology: Mononuclear Cell Isolation Protocols: T-Cell Isolation Protocols

Protocols utilize T hybridomas to detect expression of peptide-MHC complexes, since these cells provide the most convenient, consistent, and flexible T cell readout systems for these purposes. If desired, antigen-specific T cell clones can be used in lieu of T hybridoma cells, but T cell clones often give poorer responses than T hybridomas to fixed APCs due to fixation-induced loss of costimulator function. - [Read Presenting Exogenous Antigen to T Cells Protocol]

Category: Immunology: B Cell Protocols

Describes procedures for measuring the capacity of purified B cells to undergo proliferation. The method centers on the use of polyclonal stimulating agents (mitogens) because these agents stimulate the majority of B cells and because the alternative (measurement of antigen-induced proliferation) requires the laborious procedures of isolating antigen-specific B cells (which are otherwise present in too low a concentration in whole B cell populations). - [Read Proliferative Assays for B Cell Function Protocol]

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]

Category: Transfection Protocols: Stable Transfection Protocols

Stably transfected cells, generated in the first two stages of the procedure, are induced for expression of the target gene. After harvesting and lysis, the lysates are analyzed by SDS-PAGE and immunoblotting. - [Read Tetracycline as Regulator of Inducible Gene Expression III]

Category: Primary Cell Isolation and Culture Protocols

Thymosin β4 (Tβ4)-induced outgrowth and differentiation of vasculogenic precursor cells. - [Read Thymosin β4 (Tβ4)-induced Outgrowth and Differentiation of Vasculogenic Precursor Cells]

Category: Mouse Protocols: Mouse Cell Culture Protocols

Protocol for thymosin β4 (Tβ4)-induced outgrowth and differentiation of vasculogenic precursor cells from adult epicardium. Includes: Preparation of EPDCs; Characterisation of EPDC Phenotypes by Immunofluorescence. - [Read Thymosin β4 (Tβ4)-induced Outgrowth and Differentiation of Vasculogenic Precursor Cells Protocol]