determining Protocols

Category: Fungi Protocols

Serum concentrations of itraconazole should be measured in patients receiving this drug to ensure that therapeutic concentrations are being achieved. This is necessary as drug absorption can be variable, and levels may be lowered by interactions with other drugs. The assay will give an indication of whether suitable blood levels have been achieved. - [Read Bioassay for Determining Itraconazole Levels in Blood]

Category: Cell Biology and Cell Culture

OptiPrep Application sheet C14 describes procedures for determining the density and sedimenting properties of any cell (of any size or density) using either a continuous or discontinuous gradient of iodixanol. This Application Sheet describes procedures aimed at isolating specifically a relatively low-density cell fraction from any tissue. - [Read C25 Isolation from spleen, thymus, pancreas, alveolar tissue and other tissues]

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]

Category: Immunology: Immunoprecipitation Protocols: Chromatin Immunoprecipitation Protocols

Chromatin immunoprecipitation is a method for determining whether a particular protein binds to a particular DNA sequence in vivo. - [Read Chromatin Immunoprecipitation in Yeast Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Cycle Analysis Protocols

Describes assays used to determine the distribution of a population of cells to the different stages of the cell cycle as analyzed by flow cytometry. Staining the DNA with different fluorescent dyes, propidium iodide or DAPI, is one of the most direct ways of staging the cells based on DNA content. - [Read Determining Cell Cycle Stages by Flow Cytometry Protocol]

Category: Cell Biology and Cell Culture: Cell Viability Protocols

Cell-based assays are important tools for contemporary biology and drug discovery because of their predictive potential for in vivo applications.This assay gives ratiometric, inversely proportional values of viability and cytotoxicity (Figure 4.15) that are useful for normalizing data to cell number. Also, this reagent is compatible with additional fluorescent and luminescent chemistries. - [Read Determining Number of Live and Dead Cells in Cell Population: Cytotoxicity Assay Protocol]

Category: Yeast Cell Culture Protocols: Yeast Sporulation Protocols

Direct method for determining efficiency in yeast. The plating efficiency of a strain is a measure of the percentage of cells in a culture that are capable of forming colonies (colony forming . - [Read Determining Plating Efficiency in Yeast: Direct Method]

Category: Yeast Cell Culture Protocols: Yeast Sporulation Protocols

Protocol for the indirect method of determining plating efficiency in yeast. - [Read Determining Plating Efficiency in Yeast: Indirect Method Protocol]

Category: Microscopy Protocols: Fluorescence Microscopy Protocols

Most biological specimens are relatively transparent, so details of internal and intracellular morphology are difficult to image in untreated living specimens using simple bright-field techniques. Fluorescence microscopy offers greater advantages and possibilities for increasing contrast and determining the specific localization of molecules in cells. Article outlines the three methods most commonly used to introduce an appropriate label into Drosophila tissue without perturbing the process. - [Read Fluorescent Reagents for Live Cell Imaging and Their Introduction into Cells]

Category: DNA Protocols: SNP Protocols

High-resolution SNP mapping by denaturing HPLC. A SNP mapping procedure that relies on resolving polymorphisms by denaturing HPLC without the necessity of determining the nature of the SNPs. They demonstrate the use of denaturing high-performance liquid chromatography to identify mutations in the candidate genes and to fine-map chromosomal breakpoints. - [Read High-resolution SNP mapping by denaturing HPLC]

Category: Fungi Protocols: Neurospora Protocols

Information on how to use fluffy testers for determining mating type and for other applications. - [Read How to Use Fluffy Testers for Determining Mating Type and for Other Applications]

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to use helper strains for maintaining and crossing handicapped recessive mutants, for forcing and resolving heterokaryons, and for determining heterokaryon compatibility. - [Read How to Use Helper Strains for Maintaining and Crossing Handicapped Recessive Mutants]

Category: Microscopy Protocols: Electron Microscopy Protocols

The scanning transmission electron microscope precision and reproducibility of mass measurements are comparable with those of the analytical ultracentrifuge, the possibility of determining the mass not only of entire supramolecular assemblies but also of their distinct components has opened exciting new avenues which have occasionally been entered but are not yet fully explored. Includes: Principle and application (The GroEL:GroES complex). - [Read Imaging and Measuring Biomolecules & Their Assemblies by Scanning Transmission Electron Microscopy]

Category: Microscopy Protocols

The atomic force microscope (AFM) is one of the most powerful tools for determining the surface topography of native biomolecules at subnanometer resolution. The AFM can also provide insight into the binding properties of biological systems. In order to determine the specific interaction between two kinds of molecules (e.g., avidin and biotin). Includes information on principle of AFM and application of AFM. - [Read Imaging, Measuring and Manipulating Native Biomolecular Systems with the Atomic Force Microscope]

Category: Yeast Protocols

Simple method for determining the cell density of yeast cultures. - [Read Measuring Yeast Cell Density by Spectrophotometry Protocol]

Category: Cell Biology and Cell Culture: Cell Viability Protocols

The MTT Cell Proliferation Assay measures the cell proliferation rate and conversely, when metabolic events lead to apoptosis or necrosis, the reduction in cell viability. The number of assay steps has been minimized as much as possible to expedite sample processing. The MTT Reagent yields low background absorbance values in the absence of cells. Includes: Determining optimal cell counts, performing an assay, data interpretations and troubleshooting. - [Read MTT Cell Proliferation Assay Protocol]

Category: Peptide Protocols: Dissolving Handling Peptides

Peptide Handling Guides. Storage Guidelines for Lyophilized Peptides, Strategy for Dissolving Single Peptides, Determining Solubility Characteristics of peptides, Dissolving Approach for Charged Peptides, Dissolving Approach for Hydrophobic/Uncharged Peptides, Guidelines for Dissolving Several Peptides, Peptide Stability and Potential Degradation Pathways, and storage of peptides. Sigma Aldrich. PDF. - [Read Peptide Handling Guides]

Category: Fungi Protocols

Serum concentrations of voriconazole should be measured in patients receiving this drug to ensure that therapeutic levels are being achieved. The assay will give an indication of whether suitable blood levels have been achieved. - [Read Protocol: Bioassay for Determining Voriconazole Levels in Blood]

Category: PCR Protocols: Reverse Transcriptase RT-PCR Protocols

In the first part of this protocol, the linear range of amplification is determined by carrying out 10 identical PCRs in the presence of [{alpha}-32P]dCTP and stopping one reaction after every two cycles.  Amplification products are quantified on a denaturing polyacrylamide gel and the results plotted on a graph (counts per minute vs. cycle number).  Total RNA is used as an internal control. - [Read Relative RT-PCR: Determining the Linear Range of Amplification and Optimizing the Primers:Competimer]

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

In vitro transcription systems includes instructions for use of products P1420, P1430, P1440, P1450 and P1460.Includes information and protocols on RNA Transcription in vitro. Information on DNA Template Preparation;Synthesis of High-Specific-Activity Radio labeled RNA Probes;Determining Percent Incorporation and Probe Specific Activity;Removal of the DNA Template Following Transcription;Removal of unincorporated nucleotides;Synthesis of large amounts of RNA;Capping RNA for in vitro translation. - [Read Riboprobe In Vitro Transcription Systems]

Category: Cytoskeleton Protocols: Microtubules Protocols

Protocol for tubulin polymerization with GTP/GMPCPP/ Taxol. Includes: Solutions & Supplies; Prepolymerization Clarification; GTP Polymerization; Taxol Polymerization; GMPCPP Polymerization; Determining Concentration of GMPCPP/Taxol MTs. - [Read Tubulin Polymerization with GTP/GMPCPP/Taxol Protocol]

Category: General Research Protocols and Methods: Spectrophotometry Protocols

Determining the concentration of a RNA or DNA sample by using UV spectrophotometry. Hofstra Univ. Beverly Clendening. - [Read UV Spectrophotometric Analysis of DNA and RNA]