sensitivity Protocols

Category: Yeast Protocols: Yeast Genetics Protocols

Protocol describes an assay where it requires growing saturated cultures of yeast, counting, and spotting serial dilutions of yeast on both CSM and CSM + 6AU plates. - [Read 6-Azauracil Sensitivity Assay for Yeast Protocol]

Category: Drosophila Protocols

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]

Category: Signalling Signal Transduction Protocols

Protocol allows you to measure the content of cyclic adenosine 3',5'-monophosphate (cyclic AMP or cAMP) in splenic B lymphocytes (B cells) in an enzyme-linked immunoassay. This protocol utilizes acetylation of cAMP to improve sensitivity and reduce interference. Protocol includes information on: how to determine cAMP, calculations and reagents and materials. - [Read Assay of Cyclic AMP in Lysates of Cells]

Category: Cytogenetics Protocols

Protocol for comparative genomic hybridization (CGH). Includes: Theory of CGH; Sensitivity; Applications of CGH. - [Read Comparative Genomic Hybridization (CGH) Protocol]

Category: Genetics and Genomics: Cancer Genetics Protocols

Detailed, step-by-step protocol of the MethyLight assay for detection of CIMP with high sensitivity and specificity in colorectal cancer using a five marker panel composed of CACNA1G, IGF2, NEUROG1, RUNX3 and SOCS1. - [Read Determination of the CpG Island Methylator Phenotype (CIMP) in Colorectal Cancer using MethyLight]

Category: Nucleic Acid Modification Protocols: DNA Methylation Protocols

Determination of the CpG Island Methylator Phenotype (CIMP) in colorectal cancer using MethyLight. Protocol describes a detailed, step-by-step protocol of the MethyLight assay for detection of CIMP with high sensitivity and specificity in colorectal cancer using a five marker panel composed of CACNA1G, IGF2, NEUROG1, RUNX3 and SOCS1. - [Read Determination of the CpG Island Methylator Phenotype (CIMP) in colorectal cancer using MethyLight]

Category: PCR Protocols: Real-Time PCR Protocols

Protocol provides a detailed, step-by-step protocol of the MethyLight assay for detection of CIMP with high sensitivity and specificity in colorectal cancer using a five marker panel composed of CACNA1G, IGF2, NEUROG1, RUNX3 and SOCS1. - [Read Determination of the CpG Island Methylator Phenotype (CIMP) in colorectal cancer using MethyLight]

Category: Cytogenetics Protocols: DIG Biotin Labeled Probe Protocols

Protocol describes here a high sensitivity indirect detection procedure for DIG-labeled hybridization probes. The procedure uses the components of the HNPP Fluorescent Detection Set to form a fluorescent precipitate of HNPP (2-hydroxy-3-naphthoic acid-2’-phenylanilide phosphate) and Fast Red TR at the site of hybridization. Includes: In situ hybridization with DIG-labeled probes; Detection of DIG-labeled probes; Fluorescence microscopy. - [Read DNA In Situ Hybridization with an Alkaline Phosphatase-Based Fluorescent Detection System]

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

Protocol describes a high sensitivity indirect detection procedure for DIG-labeled hybridization probes.  The procedure uses the components of the HNPP Fluorescent Detection Set to form a fluorescent precipitate of HNPP (2-hydroxy-3-naphthoic acid-2’-phenylanilide phosphate) and Fast Red TR at the site of hybridization.  This procedure can be used to detect single copy sequences as small as 1 kb on human metaphase chromosomes. 
    - [Read DNA In Situ Hybridization with an Alkaline Phosphatase-Based Fluorescent Detection System Protocol]

Category: PCR Protocols: PCR Optimization

Method for amplifying DNA enzymatically by the polymerase chain reaction (PCR), including procedures to quickly determine conditions for successful amplification of the sequence and primer sets of interest, and to optimize for specificity, sensitivity, and yield.  The first step of PCR simply entails mixing template DNA, two appropriate oligonucleotide primers, Taq or other thermostable DNA polymerases, deoxyribonucleoside triphosphates (dNTPs), and a buffer. - [Read Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol for estimating the number of CD38 molecules on the CD8+ T lymphocytes of HIV-infected individuals. Includes: RECOMMENDATION OF VENDOR FOR PE-CD38 AND PE-CD4; VALIDATION OF LOGARITHMIC AMPLIFIER LINEARITY AND SENSITIVITY; CONSERVATION OF THE LEVEL OF CD4 ANTIGEN EXPRESSION ON CD4+ LYMPHOCYTES AND ITS USE AS A BIOLOGIC STANDARD FOR FLOW CYTOMETER INSTRUMENT CHARACTERIZATION; DETERMINATION OF THE NUMBER OF CD38 MOLECULES PER CD8+ CELL; etc.. - [Read Estimating the Number of CD38 Molecules on the CD8+ T Lymphocytes of HIV-Infected Individuals]

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to use spot tests to determine mutagen sensitivity. - [Read How to Use Spot Tests to Determine Mutagen Sensitivity]

Category: Microscopy Protocols: Electron Microscopy Protocols

In recent years, the increased sensitivity of electron detectors and the availability of low-vacuum or variable-pressure systems have allowed imaging of fresh tissue samples without the need for fixation, drying, and coating. This obviously saves a lot of time, although the image quality may not be as good as that obtained from fixed samples. However, for most applications that tend to be at a relatively low magnification, the quality can be as good as that obtained from fixed samples. - [Read Imaging of Fresh Arabidopsis Tissues in the Scanning Electron Microscope]

Category: Plant Biology Protocols

In recent years, the increased sensitivity of electron detectors and the availability of low-vacuum or variable-pressure systems have allowed imaging of fresh tissue samples without the need for fixation, drying, and coating. This obviously saves a lot of time, although the image quality may not be as good as that obtained from fixed samples. However, for most applications that tend to be at a relatively low magnification, the quality can be as good as that obtained from fixed samples. - [Read Imaging of Fresh Arabidopsis Tissues in the Scanning Electron Microscope Protocol]

Category: Protein Protocols: Protein Quantitation Concentration Protocols

Find a list of assays for the determination of protein concentration in a solution.  This list includes the sensitivity range, volume/amount of sample needed, subjective comments on accuracy and convenience, and major interfering agents.  Procedural details, equipment requirements, and references are outlined in the individual assay documents. - [Read List of Protein Assays]

Category: Nucleic Acid Detection Protocols

Protocol for potassium permanganate sensitivity assay. - [Read Potassium Permanganate Sensitivity Assay]

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]

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: Protein Protocols: Protein Trafficking

Methods on protein trafficking. Information on N5 Secretion and protein trafficking. Methods include: Alcian Blue Test; Lucifer Yellow uptake assay; Preparation of total protein extracts for western immunoblots; Screen for Drug Sensitivity. - [Read Protein Trafficking Methods]

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: Western Blot Protocols: Antigen Detection Methods Protocols

Use of the chemiluminescence-producing alkaline phosphatase substrate 3-(4-methoxyspiro[1,2-dioxetane-3,2'-tricyclo-[3.3.1.1(3,7)]decan]-4-yl)phenyl phosphate (AMPPD, also known as adamantyl-1,2-dioxetane phosphate), or its dioxetane relatives provides a substantial increase in sensitivity over colorimetric substrates and radiochemical methods currently used for the detection of antigen-antibody complexes immobilized on nylon or PVDF membranes. - [Read Western Analysis Using the Chemiluminescent Alkaline Phosphatase Substrate CSPD Protocol]