detection Protocols

Category: Antibody Protocols: Antibody Labeling Protocols

This protocol describes a simple chemical oxidation method for labeling antibodies with iodine. Iodide-125 (supplied as NaI) is oxidized to form iodine-125 (I2), which attacks tyrosyl and histidyl side chains. The iodinated antibodies are easily detected and quantitated using gamma counters or film. They are used primarily in immunoassays, but other techniques can be adapted conveniently to the iodine detection method. - [Read Labeling Antibodies with Iodine Protocol]

Category: Transgenic Mouse Protocols: Mouse Tail DNA Extraction Genotyping

ROSA26 : Lac-Z Detection in Tail Biopsies, the Jackson Laboratory. - [Read Lac-Z Detection in Tail Biopsies]

Category: Cell Biology and Cell Culture: Apoptosis Protocols

Detection of Fragmented DNA in Apoptotic Cells Embedded in LR White:Histochemical (LM) and Ultrastructural (EM). Goping et al 1999. - [Read Light and Electron Microscopy Detection of Apoptosis]

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

Protocol for measuring apoptosis and necrosis by dual laser flow cytometry. Includes: Preparation of stock solutions of HO342, PI, and 7-AAD; Staining for detection of apoptosis; - [Read Measuring Apoptosis and Necrosis by Dual Laser Flow Cytometry Protocol]

Category: Western Blot Protocols: Antigen Detection Methods Protocols

Protocol describes, samples containing the target protein are deposited onto a polyvinyldifluoride (PVDF) membrane using a vacuum manifold.  The immobilized protein is exposed to an antibody specific for the target protein, followed by an antibody that reacts with species-specific determinants carried by the primary antibody and is conjugated to horseradish peroxidase (HRP). - [Read Measuring Protein Concentration by Western Analysis Using Enhanced Chemiluminescence Detection]

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

Exponentially growing cells are asynchronous with respect to the cell cycle stage. Detection of cell cycle-related events is improved by enriching the culture for cells at the stage during which the particular event occurs. Methods for synchronizing cells are provided here, including those based on morphological features of the cell. - [Read Methods for Synchronizing Cells at Specific Stages of the Cell Cycle]

Category: Bacterial Protocols

Ice tea has a complex composition, which leads to reduced filterability, and a decrease in sample throughput. Its composition can generate background or false positive signals. It is also well known that ice tea contains molecules that can inhibit the bioluminescence reaction, which can generate false negative results. The aim of this study was to develop a protocol that was able to neutralize these affects and enable faster detection of contamination. - [Read Microbial Detection in Ice Tea Using the Millipore Milliflex Rapid Microbiology Detection System]

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 Organelle Protocols: Mitochondria Protocols

Kit for detecting the mitochondrial membrane potential. - [Read Mitochondrial Membrane Potential Detection Kit]

Category: Plant Biology Protocols

The standard protocol for in situ hybridizations in plants still involves fixing fresh tissue, embedding the tissue in wax, sectioning with a microtome and detection of the transcripts of interest using labeled RNA-probes. This protocol concentrates only on nonradioactive methods, as they are easy to perform, very sensitive and even faster than techniques involving radioisotope labels. - [Read Molecular and Biochemical Analysis of Arabidopsis Protocol]

Category: Cell Culture Protocols: Cell Fixing Protocols

Mounting media for immunohistology must be compatible with the detection method used. A suitable non-aqueous mounting medium is DPX. - [Read Mounting Samples in DPX Protocol]

Category: Cell Culture Protocols: Cell Fixing Protocols

Mounting media for immunohistology must be compatible with the detection method used. Suitable aqueous media are made from Gelvatol or Mowiol. - [Read Mounting Samples in Gelvatol or Mowiol Protocol]

Category: Immunology: T Cell Protocols

Mouse Protocol: Stimulation of mouse peripheral T cells with plate-bound 145-2C11 monoclonal antibody; MTT assay for detection of cellular proliferation. - [Read MTT Assay for Detection of Cellular Proliferation]

Category: Cytogenetics Protocols: In Situ Hybridization Protocols

Protocol for multiple-target DNA in situ hybridization with enzyme-based cytochemical detection systems. Includes: Cell preparations; Cell processing; Probe preparation; Multiple-target in situ hybridization (ISH); ISH with separate probe and target denaturation [for probes with repetitive (e.g., Alu) elements]; Post-hybridization washes; Enzyme-based cytochemical detection; etc.. - [Read Multiple-Target DNA In Situ Hybridization with Enzyme-Based Cytochemical Detection Systems Protocol]

Category: Genetics and Genomics: Genotyping Protocols: Mutation Detection Protocols

Protocol for mutation detection by single-strand conformational polymorphism (SSCP). Includes: PCR Protocol; SSCP Gels; - [Read Mutation Detection By Single-Strand Conformational Polymorphism (SSCP) Protocol]

Category: Genetics and Genomics: Cancer Genetics Protocols: Single Strand Conformational Polymorphism(SSCP) Protocols

Protocol for mutation detection by single-strand conformational polymorphism (SSCP). Includes: PCR Protocol; SSCP Gels. - [Read Mutation Detection By Single-Strand Conformational Polymorphism (SSCP) Protocol]

Category: Genetics and Genomics: Genotyping Protocols: Mutation Detection Protocols

This protocol is the first of two describing solid-phase chemical cleavage of mismatch (SpCCM), a method for the detection of mutations in genomic DNA. DNA fragments up to 2 kb in length can be analyzed to determine the position (within 10-15 bp) and identity of the mismatched nucleotide. - [Read Mutation Detection by Solid-Phase Chemical Cleavage of Mismatches Using Radioactive Probes Protocol]

Category: Cell Culture Protocols: Cell Culture Contamination Prevention and Removal Protocols

One of the most important, but frequently overlooked, cell culture procedures is testing cultures for microbial contamination, especially mycoplasma. It is critical for every cell culture laboratory to only use cell lines that have been carefully screened for mycoplasma. Fortunately, there is a simple fluorochrome DNA staining test that can detect both mycoplasma and virtually any other prokaryote contaminants. - [Read Mycoplasma Detection Using DNA Staining Protocol]

Category: Nucleic Acid Modification Protocols: DIG Biotin Labeled Probes Protocols

Protocol for nonradioactive in situ hybridization to polytene chromosomes with a DIG-labeled DNA probe. Includes: Labeling the hybridization probe; Preparation and denaturation of polytene chromosomes from Drosophila, Chironomus, or other species; Hybridization and detection. - [Read Nonradioactive In Situ Hybridization to Polytene Chromosomes With a DIG-Labeled DNA Probe Protocol]

Category: Nucleic Acid Detection Protocols

Seminar for nonradioactive labeling and detection. - [Read Nonradioactive Labeling and Detection Seminar]

Category: Nucleic Acid Detection Protocols: Nucleic Acid Blotting Protocols

The protocol consists of a method for the generation of cytoplasmic extracts from mammalian cells (in this case, 293T cells) without the disruption of polyribosomes, the separation of ribosomal components and polyribosomes by sucrose gradient centrifugation, the isolation of mRNA from these fractions, and detection of mRNA by Northern blot analysis. - [Read Northern Blot Analysis of mRNA From Mammalian Polyribosomes Protocol]

Category: Fungi Protocols: Neurospora Protocols

Northern blot protocol for the detection of RNA in Neurospora. Includes: Extract RNA from tissue powder; Electropheresis and transfer RNA; Preparing the membrane for probing and preparation of a riboprob.; Hybridization. - [Read Northern blot Protocol for the Detection of RNA in Neurospora]

Category: PCR Protocols: In-situ PCR Protocols

PCR IN SITU: NEW TECHNOLOGIES WITH SINGLE CELL RESOLUTION FOR THE DETECTION AND INVESTIGATION OF VIRAL LATENCY AND PERSISTENCE. Staskus 1998 U.Minn. - [Read PCR IN SITU: NEW TECHNOLOGIES WITH SINGLE CELL RESOLUTION]

Category: Genetics and Genomics: Genotyping Protocols: PCR Genotyping

Using molecular marker technology in studies on plant genetic diversity. DNA-based technologies: PCR-based technologies Amplified fragment length polymorphisms (AFLPs. Includes: AFLP technology, step by step; DNA digestion and ligation; PCRs and detection; Summarising the technology. - [Read PCR-Based Technologies Amplified Fragment Length Polymorphisms (AFLPs)]

Category: Plant Biology Protocols: Arabidopsis Tissue Culture

It is desirable to prepare subcellular fractions, either to localize proteins or to improve the sensitivity of protein detection. This procedure describes the enrichment of chloroplasts from Arabidopsis. - [Read Preparation of Arabidopsis Chloroplasts Protocol]