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Cell Viability Assays that Measure Metabolic Capacity Protocol - http://www.promega.com/paguide/chap4.htm#title3

Category: Cell Biology and Cell Culture: Cell Viability Protocols

The CellTiter-Blue® Cell Viability Assay uses an optimized reagent containing resazurin. The homogeneous procedure involves adding the reagent directly to cells in culture at a recommended ratio of 20µl of reagent to 100µl of culture medium. - [Read Cell Viability Assays that Measure Metabolic Capacity Protocol]

Cell Viability Information For Protocols and Applications - http://www.promega.com/paguide/chap4.htm

Category: Cell Biology and Cell Culture: Cell Viability Protocols

Choosing a cell viability or cytotoxicity assay from among the many different options available can be a challenging task. Includes information on: Establishing an In Vitro Model System; Choosing an Endpoint to Measure; Characterizing Assay Responsiveness; Determining Dose and Duration of Exposure; Homogeneous Assays for Multiwell Formats and Automated Screening; Additional Factors to Consider When Choosing a Cell Viability Assay; Cell Viability Assays that Measure ATP Protocol; etc.. - [Read Cell Viability Information For Protocols and Applications]

CellTiter-Glo Luminescent Cell Viability Assay Protocol - http://www.promega.com/tbs/tb288/tb288.pdf

Category: Cell Biology and Cell Culture: Cell Viability Protocols

Protocol for cell titer- glo luminescent cell viability assay. - [Read CellTiter-Glo Luminescent Cell Viability Assay Protocol]

CellTiter-Glo Luminescent Cell Viability Assay Protocol - http://www.promega.com/tbs/tb288/tb288.pdf

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

Protocol for CellTiter-Glo luminescent cell viability assay. This assay is a homogeneous method of determining the number of viable cells in culture based on quantitation of the ATP present, which signals the presence of metabolically active cells. - [Read CellTiter-Glo Luminescent Cell Viability Assay Protocol]

Chambers for Examination of Live Cells under Mechanical Stress Protocol - http://www.cshprotocols.org/cgi/content/extract/2007/3/pdb.ip32

Category: Microscopy Protocols: Live-Cell Imaging Protocols

Manual measurement and manipulation of the cell surface requires access to the cells, usually in an open chamber. Temperature-controlled chambers or stage inserts are preferred for maintaining physiological activity during the experiment. For example, heated culture dishes with coverslip glass bottoms (Bioptechs) permit high-resolution fluorescence microscopy of living cells during force application. - [Read Chambers for Examination of Live Cells under Mechanical Stress Protocol]

Characterization of Cells by Flow Cytometry Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000018.pdf?pid=PP00000018

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol uses specific antibodies coupled to one of four fluorochromes: fluorescein isothiocyanate (FITC), R-phycoerythrin (PE), peridinin chlorophyll-a (PcP), and allophycocyanin (APC). These fluorochromes can be used simultaneously to stain and analyze the expression patterns of four different proteins in the same sample. The fluorochrome stained cell populations are analyzed using a FACSCalibur dual-laser flow cytometer. - [Read Characterization of Cells by Flow Cytometry Protocol]

Characterization of Cells by Flow Cytometry Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000018.pdf?pid=PP00000018

Category: Immunology: B Cell Protocols

Fluorochromes can be used simultaneously to stain and analyze the expression patterns of four different proteins in the same sample. The fluorochrome stained cell populations are analyzed using a FACSCalibur dual-laser flow cytometer. - [Read Characterization of Cells by Flow Cytometry Protocol]

Chemotaxis Assay - http://www.cbrinstitute.org/labs/springer/protocols/melissa_chemotaxis.html

Category: Immunology: Chemotaxis

Chemotaxis Assay, Springer Lab. A chemotaxis assay's function is to assess whether a factor or molecule of interest has chemotactic activity on a motile cell type. Chemotaxis is the ability of a factor to cause the migration of a cell. The chemotactic assay is based on the creation of a chemical gradient of the chemotactic agent which will cause cells to migrate through the gradient towards the chemotactic agent. - [Read Chemotaxis Assay]

Chemotaxis Assay Protocol - http://www.cbrinstitute.org/labs/springer/protocols/melissa_chemotaxis.html

Category: Cell Biology and Cell Culture

This chemotaxis assay protocol is based on the premise of creating a gradient of the chemotactic agent and allowing cells to migrate through a membrane towards the chemotactic agent. A chemotaxis assay can determine whether your protein or small molecule of interest has chemotactic activity on a specific cell type. Chemotaxis is then the ability of a protein to direct the migration of a specific cell. - [Read Chemotaxis Assay Protocol]

Chemotaxis Practical - http://dictybase.org/techniques/chemotaxis/Soldati-Chemotaxis_Protocol.pdf

Category: Immunology: Chemotaxis

Chemotaxis Practical. Thierry Soldati. Department of Biological Sciences Animal and Plant Physiology. The assay in brief. Safety and Good Laboratory Practice: Working in a Cell Biology Laboratory. I-Chemotaxis Assay. Observation of Dictyostelium development stages. - [Read Chemotaxis Practical]

ChIP ping for Dummies - http://mcardle.oncology.wisc.edu/sugden/Protocols/html%20files/ChIPping%20for%20Dummies.htm

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

Very nice and detailed ChIP assay protocol procedure. Blocking Staph A cells, Crosslinking cells, Chromatin preparation from cell pellets, sonication, blocking. Sugden Laboratory - [Read ChIP ping for Dummies]

Chromatin Immunoprecipitation and Microarray-Based Analysis of Protein Location Protocol - http://www.nature.com/nprot/journal/v1/n2/abs/nprot.2006.98.html

Category: Immunology: Immunoprecipitation Protocols: Chromatin Immunoprecipitation Protocols

Genome-wide location analysis, also known as ChIP-Chip, combines chromatin immunoprecipitation and DNA microarray analysis to identify protein-DNA interactions that occur in living cells. Protein-DNA interactions are captured in vivo by chemical crosslinking. Cell lysis, DNA fragmentation and immunoaffinity purification of the desired protein will co-purify DNA fragments that are associated with that protein. - [Read Chromatin Immunoprecipitation and Microarray-Based Analysis of Protein Location Protocol]

Chromatin Immunoprecipitation Protocol - http://www.microarrays.ca/support/PDFs/ChIP_Protocol_for_Microarray_Analysis_Staph_A_Cells_09-07-04.pdf

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

ChIP Staph A cell preparation, preblocking and ChIP IP preclear, recovery, wash steps based on Farnham Lab Protocols. - [Read Chromatin Immunoprecipitation Protocol]

Clonality - X Chromosome Inactivation Assay Protocol - http://cgap-mf.nih.gov/Protocols/DNARNAProteomicAnalysis/DNA/DNAProtocols/Clonality.html

Category: Genetics and Genomics: Cancer Genetics Protocols

Investigators can utilize X chromosome inactivation (methylation) to determine the clonality status of a tumor or premalignant lesion in females. The technique is based on a methylation-sensitive restriction enzyme and analysis of a polymorphic locus on the X chromosome. Clonal cell populations will show "loss" of the non-methylated allele after restriction digest. The assay can be performed on DNA recovered from microdissected samples. Both frozen tissue and fixed-embedded tissue can be used. - [Read Clonality - X Chromosome Inactivation Assay Protocol]

Collecting Two-Cell- to Compacted Morula-Stage Embryos Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4359

Category: Stem Cell Protocols: Stem Cell Injection Protocols

This protocol describes a method for collecting two-cell- to compacted morula-stage embryos. Such embryos can then be used for microinjection. - [Read Collecting Two-Cell- to Compacted Morula-Stage Embryos Protocol]

Colorimetric Cytotoxcity Assays for Anchorage Dependent Cells Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/43.htm

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

The potential cytotoxicity of compounds under hypoxic conditions is determined by exposing cell cultures to test compounds in a low oxygen atmosphere. Subsequent cell survival is determined by the MTT and methylene blue colorimetric assays. - [Read Colorimetric Cytotoxcity Assays for Anchorage Dependent Cells Protocol]

Combined DNA In Situ Hybridization and Immunocytochemistry Protocol - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_127-134.pdf

Category: Cytogenetics Protocols: In Situ Hybridization Protocols

Protocol for combined DNA in situ hybridization and immunocytochemistry for the simultaneous detection of nucleic acid sequences, proteins, and incorporated BrdU in cell preparations. Includes: Cell preparations and BrdU labeling; Detection of antigen by immunocytochemistry (ICC); Visualization of ICC antigen; -Gal-BCIG reaction (for producing a blue precipitate visible under brightfield microscopy); Cell processing for in situ hybridization; In situ hybridization (ISH); etc... - [Read Combined DNA In Situ Hybridization and Immunocytochemistry Protocol]

Competent cell preparation - http://www.chem.uga.edu/scottgrp/GrpProtocols/Competent_cell_preparation.htm

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Bacterial E.Coli competent cell preparation. Calcium Chloride Protocol, Preparation of calcium chloride competent cells for frozen storage, Electroporation Protocol, Electroporation Protocol for transformations using double-stranded plasmids, Reference: - [Read Competent cell preparation]

Competent Cell Preparation [CaCl2] - http://www.cf.ac.uk/biosi/staff/kille/Methods/Gene/Comp_cell_CaCl.html

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Nice protocol for the preparation of Bacterial cells to allow uptake of plasmid vectors. Dr Peter Kille. Hot Metals Cardiff. - [Read Competent Cell Preparation [CaCl2]]

Configuration Column Construction Column Packing for Capillary Liquid Chromatography - http://cshprotocols.cshlp.org/cgi/content/abstract/2007/3/pdb.prot4578

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol for configuration, column construction, and column packing for a capillary liquid chromatography system. Protocol describes a procedure for adapting conventional HPLC systems to provide accurate low-flow rates (0.4-4 µl/min) and gradients required to operate slurry-packed capillary columns. A key component of this system is a commercial axial-beam longitudinal flow cell that can be fitted to several commercial UV detectors. - [Read Configuration Column Construction Column Packing for Capillary Liquid Chromatography]

Constructing and Expressing GFP Fusion Proteins - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4649

Category: Reporter Gene Assays: GFP Assay Protocols

The following protocol can be used for the development of stable cell lines expressing GFP fusion proteins. Although optimal transfection procedures (e.g., calcium phosphate, electroporation, or FuGENE 6 [Roche Applied Science]) vary depending on cell type, this general transfection procedure has been successful for stable transfection of HeLa, A-431, U2OS, BHK, and HT1080 cells. - [Read Constructing and Expressing GFP Fusion Proteins]

COS cell protocols - http://webusers.physics.umn.edu/~josephs/cell.html

Category: Cell Biology and Cell Culture: Cell Line Culture Conditions

COS cell protocols. Joe Skinner - [Read COS cell protocols]

COS Cell Transfection Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/C5.html

Category: Transfection Protocols

COS cell transfection protocol. Includes solutions: DEAE Dextran and 1000X Chloroquine - [Read COS Cell Transfection Protocol]

Cotransfection of Luciferase Reporter Plasmids with siRNA Duplexes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4342

Category: Reporter Gene Assays: Luciferase Assay Protocols

It is possible that some cell lines have lost the ability to perform RNAi or that cells derived from certain tissues do not support RNAi. This reporter assay, for RNAi in mammalian cells, can be used to establish whether the cells under study are susceptible to RNAi. - [Read Cotransfection of Luciferase Reporter Plasmids with siRNA Duplexes Protocol]

Counterstains Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4338

Category: Cell Culture Protocols: Cell Staining Protocols

Counterstains are used to help differentiate the various cell types of subcellular structures seen in cell staining. They are essential for tissue sections, allowing the identification of the cell types, but also may be helpful in other staining reactions. - [Read Counterstains Protocol]

Articles (1-10 of 28)

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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