Enzyme-linked reagents give excellent sensitivity and use a simple light microscope for detection. A range of enzymes is available, but for staining in situ, horseradish peroxidase will suit most needs. Diaminobenzidine (DAB) is one of the most sensitive substrates for horseradish peroxidase. It yields an intense brown product that is insoluble in both water and alcohol. It can be made more sensitive by adding metal salts such as cobalt or nickel to the substrate solution. - [Read Antibody Addition to Drosophila Specimens and Detection Using Enzyme-Linked Reagents Protocol]
Protocol describes the use of a basic water-based dye to stain for acid mucosubstances and acetic mucins located on the cartilage of fixed embryos, permitting the examination of cartilage formation. - [Read Cartilage Staining of Xenopus tropicalis Protocol]
Concentration of DNA by Ethanol Precipitation Protocol. Adapted fromBruce A. Roe, Department of Chemistry and Biochemistry, The University of Oklahoma, Norman, Oklahoma. Usually 2.5 - 3 volumes of ethanol and/or acetate solution is added to the DNA in a microcentrifuge tube. This is then put into an ice-water bath for at least 10 minutes. The precipitation is performed by incubation at -20C overnight. - [Read Concentration of Oligo DNA by Ethanol Precipitation Protocol]
Protocol provides methods for cryofreezing and subsequent thawing of mammalian cells. Pre-confluent cells are trypsinized, pelleted, resuspended in freezing medium, and gradually frozen. When needed, frozen cells are thawed quickly under running tap water and transferred to growth medium. - [Read Cryopreservation of Mammalian Culture Cells: Preparation and Recovery of Samples Protocol]
Protocol for extraction and purification of total RNA using TRIzol OR TRI reagent. Includes: Homogenization for Cell Suspensions; Phase Separation; RNA Precipitation; RNA Wash; Redissolving the RNA; Determination of RNA Concentration and Purity; Preparation of Rnase-free water. - [Read Extraction and Purification of Total RNA using TRIzol OR TRI Reagent Protocol]
Protocols included: USE OF RADIAL ARM MAZE TASK TO TEST BASIC WORKING MEMORY; USE OF RADIAL ARM MAZE TASK TO TEST WORKING VERSUS REFERENCE MEMORY; USE OF MORRIS WATER MAZE TASK TO TEST SPATIAL MEMORY; USE OF WATER MAZE TASK FOR SPATIAL PROBE TRIAL; USE OF WATER MAZE TASK TO TEST WORKING MEMORY. - [Read Learning and Memory Protocols]
Great peptide storage information. Lerner. Peptides are provided as lyophilized product that should be stored as dry powder at -20°C to -70°C in a desiccator, if possible. After lyophilization, peptides retain significant amounts of water. Peptides are slowly degraded particularly the cysteine-containing peptides are oxidized over time at -20°C. - [Read Peptide Storage Information]
Comparison among residential patch transition types and landscape water use efficiency, photosynthesis measurement protocol. - [Read Photosynthesis Measurement Protocol]
The water soluble, DNA intercalator, propidium iodide (PI), is used to bind to DNA after permeabilization of cells with NP40. The amount of dye bound correlates with the content of DNA within a given cell. Once cells are stained, they are analyzed on a flow cytometer. The relative content of DNA indicates the distribution of a population of cells throughout the cell cycle. - [Read Quantification of Apoptosis and Cell Cycle Distribution of Primary B Cells Using Propidium Iodide]
The water soluble, DNA intercalator, propidium iodide (PI), is used to bind to DNA after permeabilization of cells with NP40. The amount of dye bound correlates with the content of DNA within a given cell. Once cells are stained, they are analyzed on a flow cytometer. The relative content of DNA indicates the distribution of a population of cells throughout the cell cycle. - [Read Quantification of Apoptosis and the Cell Cycle Distribution of Primary B Cells Using PI]
Protocol for myelin sheath. Luxol Fast Blue is the alcohol soluble counterpart of the water soluble Alcian Blue. Staining is due to lipoproteins, and the mechanism is one of an acid-base reaction with salt formation; the base of the lipoprotein replaces the base of the dye. - [Read Staining for Myelin Sheath Protocol]
Vogel's 50X salts as given in the Microbial Genetics Bulletin 13:42-43, 1956. In 750 ml. distilled water, dissolve successively with stirring at room temperature. - [Read Vogel's Minimal Media Protocol]
3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol. This protocol contains the steps for 3' end rapid amplification of mRNA by PCR. The first-strand cDNA is synthesized from total or poly(A+) RNA by priming from the poly-A tail of the mRNA using a oligo (dT) adaptor primer. The cDNA is then amplified via PCR using a gene-specific primer and an adaptor primer.