determination Protocols

Category: Protein Protocols: Protein Phosphorylation Protocols

Paper describing methods for monitoring kinase activity, investigating kinase–substrate specificity, examining phosphorylation in planta and the determination of phosphorylation sites in a protein. In addition, strategic considerations for experimental design and variables will be discussed. Scott C. Peck, the Plant Journal. - [Read Analysis of protein phosphorylation: methods and strategies PDF]

Category: DNA Protein Interactions Protocols: Electrophoretic Mobility Shift Assays EMSA Protocols

Protocols on band shift. Includes: Phosphorylation of recombinant antibodies; Labeling the antigen with a red fluorophore; Native gel electrophoresis; Band-shift assay for the determination of Kd; Band-shift assay for the determination of koff. - [Read Band Shift Protocols]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Method is for preparing chromosomes from single flower buds of A. thaliana. It does not kill the plants allowing the determination of their chromosome number throughout development. Includes: Preparations of Arabidopsis; Preparation of Chromosome; Staining Chromosomes. - [Read Chromosome Spreads from Flower Buds of Arabidopsis thaliana Protocol]

Category: Protein Protocols: Protein Quantitation Concentration Protocols

A Comparision of Different Protein Determination Methods and protocols in table format. The Wolfson Centre for Applied Structural Biology The Hebrew University of Jerusalem Dr. Mario Lebendiker. - [Read Comparison Different Protein Quantitation Determination Methods]

Category: Plant Biology Protocols: Photosynthesis and Respiration Protocols

This protocol describes how to determine the chlorophyll content of chloroplasts using a spectrophotometric assay. - [Read Determination of Chlorophyll Content in Chloroplasts]

Category: Pharmacology and Toxicology Protocols

Protocol for the determination of IC50. - [Read Determination of IC50 Protocol]

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol for determination of neopterin and biopterin by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) in rat and human plasma, cell extracts and tissue homogenates. - [Read Determination of Neopterin and Biopterin by Liquid Chromatography 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: Protein Protocols: Protein Quantitation Concentration Protocols

Determination of the Extinction Coefficient for a Protein of Unknown Concentration. The concentration can be determined for a solution of a pure protein with unknown extinction coefficient. - [Read Determination of the Extinction Coefficient for a Protein of Unknown Concentration]

Category: Fungi Protocols: Fungi Genetics Protocols

Standard operating procedure for the determination of tissue fungal burden utilizing quantitative real time polymerase chain reaction (QPCR). This standard operating procedure will provide information on how to assess fungal tissue burden of infected animals by use of a single copy (FKS) or multicopy gene (18s RNA) to assess the number of fungal cell nuclei present. - [Read Determination of Tissue Fungal Burden Utilizing Quantitative Real Time Polymerase Chain Reaction]

Category: ELISA Protocols

Protocol for immunizations and enzyme-linked immunosorbent assay (ELISA) for antibody isotype determination. - [Read ELISA for Antibody Isotype Determination 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: Protein Microarray Protocols

Experimental Procedures for Protein Microarrays. MacBeath G & Schreiber SL (2000). Printing proteins as microarrays for high-throughput function determination. Science 289: 1760-1763. - [Read Experimental Procedures for Protein Microarrays]

Category: Nucleic Acid Purification Protocols: RNA Isolation Protocols

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]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Flow cytometric determination of leukocyte surface antigens in whole blood. Quantitation of cell surface antigens in whole blood with the flow cytometer is very simple and requires:
1. Blood collection; 2. Addition of antibody; 3. Calibration of the flow cytometer; 4. Making measurements. - [Read Flow cytometric determination of leukocyte surface antigens in whole blood]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Immunofluorescence Staining Protocols

Protocol for flow cytometric determination of leukocyte surface antigens in whole blood. Includes: Blood collection; Add antibody to the blood; Make measurements. - [Read Flow Cytometric Determination of Leukocyte Surface Antigens in Whole Blood Protocol]

Category: Reporter Gene Assays

Protocol for GUS reporter gene assay. Includes: Protein isolation; Alternative method for small (<1g) quantities of tissue; GUS assays; Bradford protein concentration determination assays - [Read GUS Reporter Gene Assay Protocol]

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast PCR Protocols

To identify the YAC subclones containing both a human insert and a portion of either the left or right arm of the pYAC4 vector. Identification of these clones is necessary in order to do YAC chromosome walking, and is also useful in the determination of whether a particular YAC clone has a contiguous human insert or whether a co-cloning event has occurred. Vector arm sequences are identified using pBR322 fragments from a BamHI-PvuII double digest. - [Read Identification of End Clones in YAC Subclone Libraries 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: Protein Protocols: Lowry Protein Assay

Lowry – Protein Determination. Quantitation of total protein using the lowry protein assay. Dr. Mario Lebendiker. The Wolfson Centre for Applied Structural Biology, The Hebrew University of Jerusalem. Method materials and notes for the Lowry assay. - [Read Lowry – Protein Determination]

Category: Microinjection Protocols: Microinjection Equipment

This protocol describes the determination of useful settings with the Sutter Puller P-97 to make injection pipettes for microinjection. - [Read Making Injection Pipettes Protocol]

Category: Mouse Protocols: Mouse Embryology Protocols

Protocol describes the determination of useful settings with the Sutter Puller P-97 to make injection pipettes for microinjection. - [Read Making Injection Pipettes Protocol]

Category: Chromatography Protocols: Gas Chromatography Protocols

Protocol presents a method that allows rapid determination of the monosaccharide composition of glycans, glycoproteins, and proteoglycans that contain (or are suspected to contain) phosphorylated sugars.  (For samples that do not contain phosphorylated sugars see Monosaccharide Analysis by Methanolysis.) - [Read Monosaccharide Analysis of Phosphorylated Sugars by Methanolysis and Diazomethane Methylation]