detection Protocols

Category: Plant Biology Protocols: Arabidopsis Tissue Culture

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

Category: Fungi Protocols

Protocol for the preparation of solid tissue for Aspergillus galactomannan antigen detection by Platelia (Biorad). Technique was designed for use on human serum. However, it may also be possible to perform this method on solid tissues and organic solutions. Viscous solution and tissue specimens need to be pre-treated to achieve the extraction of the Aspergillus antigen and to get a homogeneous sample in solution. - [Read Preparation of Solid Tissue for Aspergillus Galactomannan Antigen Detection by Platelia Protocol]

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

Programmed Cell Death In Situ Detection Using Terminal deoxynucleotidyl Transferase (TdT). Gavrieli et al., 1993. NIEHS. - [Read Programmed Cell Death In Situ Detection Using Terminal deoxynucleotidyl Transferase]

Category: Signalling Signal Transduction Protocols

Detection of phosphorylated tyrosine residues can be performed using anti-P-TYR Ab and Western Analysis.Includes 2nd method,which uses phosphotyrosine in conjunction with anti-P-TYR Ab to "unlabel" potential proteins.By comparing Westerns developed with the 1st method(reveals phosphorylated protein) and the 2nd method(reveals non-specific labeling), a more accurate picture of those proteins phosphorylated on tyrosine can be seen. Includes: Protein Preparation, Electrophoresis and Transfer. - [Read Protocol for Antiphosphotyrosine Western Blot Analysis]

Category: Reporter Gene Assays: Luciferase Assay Protocols

Protocol for luciferase assay for in vitro detection. Protocol includes: Before cell lysate preparation; Cell lysate preparation; Protocol for manual luminometers; Protocol for plate reading luminometer; Preparation of protein assay reagent; Protein standards. - [Read Protocol for Luciferase Assay for In Vitro Detection]

Category: Cell Biology and Cell Culture

This protocol introduces the use of a liquid-filled wash chamber that separates unbound cells by gravity thereby eliminating uncontrolled shear forces and passage of adherent cells through a liquid/air interface. The cells are loaded with a fluorescent dye (6-carboxyfluorescein diacetate) for detection although other methods such as radioactive labels malabels may be used. This protocol is also useful for assaying molecules that promote or inhibit cell adhesion. - [Read Protocol for Measurement of Cell Adhesion Under Static Conditions]

Category: Cell Organelle Protocols: Nucleus Protocols

A simplified and efficient 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 Protocol for Nonradioactive In Situ Hybridization to Polytene Chromosomes With DIG-labeled DNA Prob]

Category: Cytogenetics Protocols: DIG Biotin Labeled Probe Protocols

The protocol given makes the method of in situ hybridization easier, faster, more reliable, and available to anyone who can operate a microscope. Includes: Labeling the hybridization probe; Preparation and denaturation of polytene chromosomes from Drosophila, Chironomus, or other species; Hybridization and detection. - [Read Protocol for Nonradioactive In Situ Hybridization to Polytene Chromosomes with a DIG-labeled DNA]

Category: Cell Biology and Cell Culture: Apoptosis Protocols

Quantitative Flow Cytometry for Apoptosis Detection. Flow Cytometry Facility, U.Illinois. - [Read Quantitative Flow Cytometry for Apoptosis]

Category: Cell Biology and Cell Culture

Quantitative Telomerase Detection Kit. Includes: QTD Real-Time PCR Assay Procedure; Quantitative Telomerase Detection Kit Description. - [Read Quantitative Telomerase Detection Kit]

Category: Cytogenetics Protocols: RNA In Situ Hybridization Protocols

RNA-RNA in situ hybridization using DIG-labeled probes: the effect of high molecular weight polyvinyl alcohol on the alkaline phosphatase indoxyl-nitroblue tetrazolium reaction. RNA-RNA in situ hybridization protocol using alkaline phosphatase-conjugated digoxigenin-(DIG-) labeled probes is presented. The addition of polyvinyl alcohol (PVA) of high molecular weight (40 – 100 kD) to the BCIP-NBT detection system enhances the alkaline phosphatase reaction and prevents... - [Read RNA-RNA In Situ Hybridization Using DIG-Labeled Probes Protocol]

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Single colour detection for biotinylated probes. Sanger Institute - [Read Single Colour Fish Analysis Method]

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

Single-strand confirmation polymorphism analysis (SSCP) is a powerful and robust method for the detection of DNA sequence changes (single-base substitutions) based on shifts in electrophoretic mobility. In this protocol, the target sequence is simultaneously labeled and amplified, then heat-denatured and resolved by non-denaturing polyacrylamide gel electrophoresis. - [Read Single-Strand Conformation Polymorphism Analysis Protocol]

Category: DNA Protocols: SNP Protocols

SNP detection with mutagenic primers. Input sequence will be searched to find changes in one nucleotide near the location of the SNP, so that mutagenic primers may be easily designed. Bikandi, J., San Millán, R., Rementeria, A., and Garaizar, J. In silico analysis of complete bacterial genomes: PCR, AFLP-PCR, and endonuclease restriction. Bioinformatics 2004 Mar 22;20(5):798-9. - [Read SNP detection with mutagenic primers]

Category: DNA Protocols: SNP Protocols

Compare restriction patterns of two sequences for SNP/Mutation detection. Online program that compares restriction enzyme digest of DNA and finds the differences between two sequences. - [Read SNP Restriction Digestion Comparison]

Category: Cell Biology and Cell Culture

Protocol for telomerase PCR ELISA. Fast and sensitive detection of telomerase activity. - [Read Telomerase PCR ELISA Protocol]

Category: ELISA Protocols

Protocol for telomerase PCR ELISA. For fast and sensitive detection of telomerase activity. - [Read Telomerase PCR ELISA Protocol]

Category: Radioactivity: Liquid Scintillation Counting

The Liquid Scintillation Counting Process. Great Review of the Process and detection procedure of the counter. Dr. Gambhir - [Read The Liquid Scintillation Counting Process]

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

Two distinct modes of cell death, apoptosis and necrosis, can be distinguished on the basis of differences in morphological, biochemical, and molecular changes occurring in the dying cells. Report the development of flow cytometric techniques that permit concomitant detection of apoptosis and cellular DNA content or BrdU content analysis. - [Read The Use of Flow Cytometry for Concomitant Detection of Apoptosis and Cell Cycle Analysis]

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Two-colour detection for combinations of biotinylated and digoxigenin-labelled probes. Sanger Institute - [Read Two-colour detection for combinations of biotinylated and digoxigenin-labelled probes]

Category: Immunohistochemistry Protocols

Protocol provides the sensitivity of enzymatic detection (HRP) in immunohistochemical procedures with the versatility and convenience of fluorescence detection. - [Read Tyramide Amplification and Synthesis Protocol]

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Protocol for tyramide signal amplification (TSA) detection. - [Read Tyramide Signal Amplification (TSA) Detection Protocol]

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

Protocol for VEGF antibody staining. Includes: Primary Antibody; Antigen Retrieval; Detection Methods; Chromogen Substrate; Counterstain. - [Read VEGF Antibody Staining Protocol]

Category: Viral Protocols

Methods and information related to viral detection and concentration in food. Includes: Nucleic acid amplification methods; Recent progress in polymerase chain reaction detection; Detection of human enteric viruses in non-shellfish food; Choice of primers for the detection of human caliciviruses in foods; Biosensors and microarray detection; Alternative confirmation methods. - [Read Viral Detection and Concentration in Food]

Category: Cytogenetics Protocols: In Situ Hybridization Tissue Sections Protocols

Protocol for visualizing miRNAs in adult skeletal muscle by hybridization in situ. Includes: Visualizing miRNAs in adult skeletal muscle by hybridization in situ; Probe preparation; Tissue preparation; Fixation; Deproteinization with Proteinase K; Acetylation of the tissue; Dehydration; Hybridization step; Post-Hybridization Washes (Step 1-6 should be performed on shaker); Signal Detection (Steps should be performed on a shaker); Post-Detection Washes. - [Read Visualizing miRNAs in Adult Skeletal Muscle by Hybridization In Situ Protocol]