assays Protocols

Category: Drosophila Protocols

Protocol for Drosophila larval chemotaxis assays. - [Read Drosophila Larval Chemotaxis Assays Protocol]

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

Macrophage cells in culture may be exposed to particulate matter, and resultant effects upon cell viability determined by vital dye exclusion and enzyme leakage assays. - [Read Dust Toxicity in Rat Alveolar Macrophage Cultures Protocol]

Category: Cell Biology and Cell Culture

Flow assays offer visualization of cell adhesion under wall shear stress. Visualization of the different events of cell adhesion can be quantified by selective image acquisition and subsequent image processing. Flow assays are suited for adhesive events which occur very rapidly in a time scale shorter than that of most static adhesion assays. Also, events subsequent to the initial events can be studied such as cell stabilization and spreading giving some insight into the kinetics of cell-cell. - [Read Dynamic Flow Assay for Cell Adhesion in a Parallel Plate Flow Chamber]

Category: Cell Biology and Cell Culture: Cell Adhesion Protocols

Flow assays allow visualization of cell adhesion under well-defined wall shear stress. Visualization of the events of cell adhesion are quantified by selective image acquisition and image processing. Events subsequent to the initial events can be studied such as cell stabilization and spreading. John T. Patton~GlycoTech Corporation, Rockville, Maryland - [Read Dynamic Flow Assay in a Parallel Plate Flow Chamber]

Category: Cytoskeleton Protocols: Microtubules Protocols

Protocol for flow cell assays with microtubules. Includes: Solutions & Supplies; VE-DIC Assays; Motility; Dynamics; Fluorescence Assays; Oxygen Scavenging; Assays. - [Read Flow Cell Assays with Microtubules: Motility/Dynamics in Fluorescence and VE-DIC Protocol]

Category: Nucleic Acid Electrophoresis Protocols: DNA Electrophoresis Protocols: DNA Electrophoresis in Polyacrylamide Gels Protocols

Protocol exploits differences in electrophoretic mobility through a nondenaturing polyacrylamide gel between a rapidly migrating target DNA and a more slowly migrating DNA-protein complex. - [Read Gel Retardation Assays for DNA-binding Proteins Protocol]

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: Reporter Gene Assays

Protocol for GUS reporter gene assay. Includes: Protein isolation; Alternative method for small (<1g) quantities of tissue; GUS assays; Bradford protein concentration determination assays - [Read GUS Reporter Gene Assay Protocol]

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Protocol for the hematopoietic colony forming cell assay. - [Read Hematopoietic Colony Forming Cell Assays Protocol]

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to do recombination assays. - [Read How to Do Recombination Assays]

Category: Protein Protocols: Protein Trafficking

Import assays into chloroplasts. Includes: Preparation of chloroplasts and lysates; Transport of lumenal proteins. - [Read Import Assays Into Chloroplasts: Integration or Transport into Thylakoids]

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

Protocol outlines the general procedure and requirements for in vitro translation of CFTR and outlines some assays using in vitro translated product. Core glycosylation of CFTR occurs in the ER. An assay for this processing step requires the addition of microsomal membranes to the basic in vitro translation mixture. This protocol takes this into account. - [Read In vitro Translation Assays for CFTR]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol for the isolation of Arabidopsis nuclei and measurement of gene transcription rates using nuclear run-on assays. Plant materials are ground in hexylene glycol-based buffers and highly enriched nuclear fractions are obtained using Percoll density gradients. Standard and small-scale protocols are presented, along with a tested method for nuclear run-on assays. The entire process may be completed within 3 days. - [Read Isolation of Arabidopsis Nuclei and Measurement of Gene Transcription Rates Protocol]

Category: Signalling Signal Transduction Protocols: MAPK Protocols

Nature protocol entailing a kinase assay. - [Read Kinase Assays]

Category: Protein Protocols: Protein Quantitation Concentration Protocols

Find a list of assays for the determination of protein concentration in a solution.  This list includes the sensitivity range, volume/amount of sample needed, subjective comments on accuracy and convenience, and major interfering agents.  Procedural details, equipment requirements, and references are outlined in the individual assay documents. - [Read List of Protein Assays]

Category: Protein Protocols: Lowry Protein Assay

Lowry Protein Assay. The Lowry procedure is one of the most venerable and widely-used protein assays, being first described in 1951 [Lowry et al., J. Biol. Chem. 193: 265-275 (1951)]. Under alkaline conditions, copper complexes with protein. When folin phenol reagent (phospho-molybdic-phosphotungstic reagent) is added, the Folin-phenol reagent binds to the protein. Bound reagent is slowly reduced and changes color from yellow to blue. P.J. Hansen, Dept. of Animal Sciences, University of Florida. - [Read Lowry Protein Assay]

Category: Reporter Gene Assays: Luciferase Assay Protocols

Protocol describing luciferase assays in OT-1 hybridoma cells. - [Read Luciferase Assays in OT-1 Hybridoma Cells Protocol]

Category: Cell Biology and Cell Culture: Apoptosis Protocols: Apoptosis Analysis

Measurement of Apoptosis and Other Forms of Cell Death. Jagan Muppidi, Melissa Porter, and Richard M. Siegel. As programmed cell death (PCD) or apoptosis has emerged as an important regulator of development and homeostasis in multicellular organisms, methods to quantify apoptosis and to distinguish it from necrosis have been developed. This unit presents a set of assays for these purposes, many of which are technically very simple and ideally suited to the study of hematopoietic cells. - [Read Measurement of Apoptosis and Other Forms of Cell Death]

Category: Microsphere Analysis Protocols

The role of microspheres in these screens is similar to their traditional role in immunoassays, namely as a solid phase to either enhance detection, separation, or both. The predominance of radioactive assays in high-throughput screening, along with the desire to find alternative means of detection, have led to research on substituting alternative fluorescent technologies. - [Read Microspheres for High-Throughput Screening Assays]

Category: Cell Biology and Cell Culture

Microtuble/ organelled motility assay using Golgi or ER membranes, 45 uM tubulin, rat liver cytosol, and 20x energy regeneration system. - [Read Microtubule/ organelle Motility Assays]

Category: Immunology: Chemotaxis

MONOCYTE AND NEUTROPHIL ISOLATION. The protocol describes the method used for isolation of monocytes and neutrophils used in both the chemotaxis assay and microbicidal assays. Pediatric AIDS Clinical Trials Group - [Read MONOCYTE AND NEUTROPHIL ISOLATION]

Category: Cell Biology and Cell Culture: Cell Viability Protocols

The latest generation of Promega cell-based assays includes luminescent and fluorescent chemistries to measure markers of cell viability, cytotoxicity and apoptosis, as well as to perform reporter analysis. Using these tools researchers can investigate how cells respond to growth factors, cytokines, hormones, mitogens, radiation, effectors, compound libraries and other signaling molecules. The protocols provided are guidelines for multiplexing cell-based assays & are intended as starting points. - [Read Multiplexing Cell Viability Assays Protocols]

Category: Protein Protocols: Protein Extraction From Tissue

Plant Protein Isolation - Rapid isolation of protein for SDS-PAGE analysis. (Essentially the same protocol as that described for GUS Assays). Stockinger Lab - [Read Plant Protein Isolation]

Category: Viral Culture and Isolation Protocols: Viral Isolation Protocols

Protocol for plaque assays for adenovirus titration. - [Read Plaque Assays for Adenovirus Titration Protocol]

Category: Antibody Protocols: Antibody Production Protocols

Protocol for polyclonal antibody production. Very useful for rapid and simple generation of antibodies for western blots, ELISA assays, and immunoprecipitation. Includes: Rabbit Immunization; Initial Preparation; Pre-bleed; Antigen Injection; Monitoring of Titer; Purification of Antibodies. - [Read Polyclonal Antibody Production Protocol]