common Protocols

Category: Animal Research Techniques

Anesthesia Analgesia of Mice, Rats, Rabbits, Guinea Pigs, Dogs, Cats, Macaques, Suggestions for monitoring anesthetized veterinary patients, Alternatives to the use of Methoxyflurane in Rodents, how to mix common anesthetic mixtures. Dr.Parker Univ.Iowa - [Read Anesthesia and Analgesia of Laboratory Animalsesthesia and Analgesia of Laboratory Animals]

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]

Category: Cell Immortalization Protocols

Information why cell immortalization is important. Also the available tools to be able to perform cell immortalization. - [Read Cell Immortalization]

Category: Buffers Media, and Solutions

Salmon Lab, University of North Carolina at Chapel Hill, Department, of Biology. - [Read Common Buffer List]

Category: Buffers Media, and Solutions

Recipes for commonly used buffer solutions. (Gerard R. Lazo) - [Read Common Buffers]

Category: Buffers Media, and Solutions: Buffer Recipes for DNA Work

Common DNA solutions.Dr. Rick Hershberger - [Read Common DNA solutions.]

Category: Plant Biology Protocols

In most natural habitats, Arabidopsis is a winter annual: Its seeds germinate in the fall, the young plants survive the winter, floral meristems emerge in the spring, and only the seeds survive the summer months. Most common laboratory varieties of Arabidopsis flower within 4 weeks of germination, and seeds can be collected after an additional 4-6 weeks. - [Read Cultivation of Arabidopsis Protocol]

Category: C. elegans Protocols: C. elegans Fixing Protocols

Common method for fixing worms is to use paraformaldehyde. This method provides a gentler fixation than the Bouin’s method, but often requires the use of collagenase. This method is particularly good for examining adult worms. - [Read Fixing Caenorhabditis elegans in Paraformaldehyde Protocol]

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

Common methods applicable to flow cytometry make it possible to: (1) identify and quantify dead or dying cells, (2) reveal a mode of cell death (apoptosis or necrosis), and (3) study mechanisms involved in cell death. Gross changes in cell morphology and chromatin condensation, which occur during apoptosis, can be detected by analysis with laser light beam scattering. - [Read Flow Cytometry of Apoptosis Protocol]

Category: Fungi Protocols: Aspergillus Protocols

Compendium of protocols for using Aspergillus nidulans in genetic, molecular, and cell biological investigations, originally written for members of my research group. It also summarizes our common growth media and nutritional supplements, many of which originally appeared elsewhere but now are difficult to locate. Includes: Growth and storage of Aspergillus nidulans conidia; Nutritional supplements for our common auxotrophies; Double mutants; Mitotic mapping - assigning genes to chromosomes; etc - [Read Fundamentals of Growth, Storage, Genetics and Microscopy of Aspergillus nidulans Protocols]

Category: Microinjection Protocols: Microinjection of DNA Protocols

This protocol describes a method for pulsed-flow microinjection using the Eppendorf FemtoJet injector and Eppendorf InjectMan; this is the most common type of pulsed-flow microinjection system currently being used. The advantage of this type of system over a controlled-flow system is that much more control is available over the injection parameters, reducing variability in injections. In addition, the system allows a diagonal insertion of the needle into the cell. - [Read Gene Delivery by Direct Injection (Microinjection) Using a Pulsed-Flow System Protocol]

Category: Cell Biology and Cell Culture: Troubleshooting Cell Culture Problems

Guide For Identifying And Correcting Common Cell Growth Problems. Corning. Surface Treatment Process, Problems Related to Technique, Problems Related to Incubators, Problems Related to Culture Media, Problem Solving Suggestions. - [Read Guide For Identifying And Correcting Common Cell Growth Problems]

Category: Cell Culture Protocols: Cell Immortalization Protocols

Cultivating animal cells in the laboratory is an indispensable technique for cell biologists. However, most normal primary cell lines, while faithfully reproducing the phenotype of their tissue of origin, do not grow indefinitely in culture. After a series of population doublings (the number of which varies by species, cell type, and culture conditions) primary cells enter a state where they no longer divide. - [Read Immortalization of Cells in Culture]

Category: Western Blot Protocols: Antigen Detection Methods Protocols

The blot is blocked to prevent nonspecific adsorption of the immunological reagents.  Antibodies are then bound to the proteins immobilized on the membrane, and the antigen is detected by labeling the antibodies with conveniently identified tags.  Common labeling methods for chemiluminescent detection include anti-immunoglobulin antibody-coupled enzymes such as horseradish peroxidase, which catalyzes the oxidation of luminol and in turn releases light. - [Read Immunoblotting: Antigen Detection Using Chemiluminescence Protocol]

Category: Protein Protocols: Immunoprecipitation Protocols: Immunoprecipitation Troubleshooting

Immunoprecipitation Troubleshooting Guide. Stressgen. Problem: Non-specific background, Specific Background, Specific antigen not detected. With Solutions to common IP problems. - [Read Immunoprecipitation Troubleshooting Guide PDF]

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

The plant transformation procedures described involve floral dip, vacuum infiltration, and spraying. They yield transformants at frequencies ranging up to several percent, with the most common frequency being 0.1%-1%. - [Read In Planta Transformation of Arabidopsis Protocols]

Category: Microscopy Protocols: Live-Cell Imaging Protocols

Several common drugs, their targets, and protocols are described for studying organelle distribution and trafficking. The drugs are readily available from general suppliers, including Sigma, Roche, and Calbiochem. - [Read Membrane Trafficking and Organelle Reagents]

Category: DNA Microarray Protocols: Troubleshooting DNA Microarray

Images present some common problems encountered during microarray production, with possible solutions. Galbraith laboratory. - [Read Microarray troubleshooting: image gallery]

Category: Cytoskeleton Protocols: Microtubules Protocols

Negative staining is a rapid, qualitative method for analyzing microtubule structure at the EM level. Because negative staining involves deposition of heavy atom stains, structural artifacts such as flattening of the cylindrical microtubule and opening up of microtubules into flat sheets are common. Negative staining is very useful because of its ease, rapidity and lack of requirement for specialized equipment other than that found in a regular EM facility. - [Read Negative Stain Electron Microscopy of Microtubules Protocol]

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

Cryopreserved PBMCs are a common specimen source for studies of immunological responses to vaccines, immunotherapies, etc. The health and viability of cells recovered post-cryopreservation are of course critical to the success and accuracy of immunological assays performed on them. This protocol standardizes PBMC isolation and cryopreservation techniques, specifically for the assessment of thawed cells by cytokine flow cytometry. - [Read Protocol for Isolation, Cryopreservation, and Thawing of PBMCs]

Category: Protein Protocols: Protein Extraction Protocols

Protocol for Protein Extraction Using Proteomics. Extraction of proteins from plant cells that are rich in compounds that interfere with the 2-Dimensional electrophoretic separation methods such as salts, organic acids, phenolics, pigments, terpenes, among others. A common protocol used in our lab for extraction proteins from plant tissues consists in the homogenization of mortar-grounded material in liquid nitrogen with an extraction buffer. - [Read Protocol for Protein Extraction Using Proteomics]

Category: Protein Protocols: Immunoprecipitation Protocols: Immunoprecipitation Troubleshooting

Troubleshoot Immunoprecipitation. Chemicon. The most common challenge with immunoprecipitation is trying to lower the number and type of background proteins that contaminate the washed immune complexes. Suggestions for decreasing background in IP. - [Read Troubleshoot Immunoprecipitation]