quantitation Protocols

Category: Lipid Protocols: Arachidonic Acid Protocols

Protocol for AA and metabolite quantitation of cell pellets post AA labeling. Includes: Extraction and TLC. - [Read AA & Metabolite Quantitation of Cell Pellets Post-AA Labeling Protocol]

Category: Lipid Protocols: Arachidonic Acid Protocols

Protocol for AA and metabolite quantitation of media post- AA labeling. - [Read AA & Metabolite Quantitation of Media Post-AA Labeling Protocol]

Category: Genetics and Genomics: Cancer Genetics Protocols

Describe the methods to identify and quantitate the specific A/E9a transcript in t(8;21) patients samples relative to the AML1-ETO transcript encoding the well known full-length 752 amino acid AML1-ETO protein (AE). Includes: RNA preparation and RT-PCR; Relative quantitation of the AE9a and the AE transcripts. - [Read An Alternatively Spliced Isoform of t(8;21) Transcript Promotes Leukemogenesis]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

Protocol for CellTiter-Glo luminescent cell viability assay. This assay is a homogeneous method of determining the number of viable cells in culture based on quantitation of the ATP present, which signals the presence of metabolically active cells. - [Read CellTiter-Glo Luminescent Cell Viability Assay 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: Nucleic Acid Purification Protocols: DNA Isolation Protocols: Genomic DNA Isolation Protocols

In this protocol, the DNA-binding capacity of Wizard MagneSil particles is used to capture and release a consistent amount of DNA (100 ng) across a wide range of samples.  At the end of the procedure, the DNA is eluted into 100 µl Elution Buffer to give a final concentration of 1 ng/µl, relieving the need for postpurification DNA quantitation. - [Read DNA IQ Isolation of Genomic DNA from Stains and Buccal Swabs 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: Nucleic Acid Detection Protocols

Protocol describes the quantitation of DNA using Hoechst 33258, a fluorescent dye that binds to double-stranded DNA.  Fluorometry is simple and more sensitive than spectrophotometry, and allows the detection of nanogram quantities of DNA.  The assay can only be used to measure the concentration of DNAs whose sizes exceed ~1 kb, as Hoechst 33258 binds poorly to smaller DNA fragments. - [Read Fluorometric Quantitation of DNA Using Hoechst 33258 Protocol]

Category: Immunology: T Cell Protocols

Describes generating CTL against some commonly used target antigens. Two methods for the quantitation of CTL activity are described based on the two pathways used bt CTL to kill target cells. In one pathway, they release lytic granules containing perforin and granzymes, leading to apoptosis and target cell lysis. In a second pathway, they trigger apoptosis via Fas/Fas ligand interactions. - [Read Induction and Measurement of Cytotoxic T Lymphocyte Activity Protocol]

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: Reporter Gene Assays: Luciferase Assay Protocols

Luciferase assay using a 24 well plate. Includes: Cell Lysis; Reagent Preparation; Luminescence measurement; Wash injector; Protein quantitation; Data Analysis. - [Read Luciferase Assay 24 Well Plate (Promega System)]

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols

Neuron-fibroblast coculture assay to demonstrate presynaptic differentiation.  Includes: Preparation of HEK293T cells, Coculture of HEK293T cells and neurons, Immunostaining, Vesicle turnover experiment, Image acquisition and quantitation. - [Read Neuron Fibroblast Coculture Assay to Demonstrate Presynaptic Differentiation]

Category: DNA Microarray Protocols

Reverse transcriptase labeling with Cy3/Cy5, Staining of total DNA on slides, construction, Laser scanning, Spot quantitation, Statistics, sample tracking. Vodkin Laboratory. U.Illinois. - [Read Protocols for microarray development in use in the Vodkin Laboratory]

Category: General Research Protocols and Methods: Spectrophotometry Protocols

Quantitation of DNA, RNA and ss Oligos. Mullin Lab, UCSF. - [Read Quantitation of DNA, RNA and ss Oligos]

Category: Immunology: T Cell Protocols

In the first protocol, IL-2-producing murine T cells are measured following stimulation by the mitogen Con A. The second protocol provides a modification for using human responder cells. The second protocol is used for estimating the proportion of cells that can generate a clone of cytotoxic effector cells when stimulated by Con A with the addition of IL-2. - [Read Quantitation of Functional T Cells by Limiting Dilution Protocols]

Category: PCR Protocols: Quantitative PCR Protocols

Protocol that uses the polymerase chain reaction (PCR) to quantitate the numbers of a particular DNA sequence, from 1 to 20,000 molecules per sample.  In addition, it helps assess the presence of contaminating sequences, which can seriously affect the outcome of the procedure. - [Read Quantitation of Rare DNAs by PCR Protocol]

Category: Real Time PCR

Real-Time Chemistry, Instrumentation, Real-Time PCR Quantitation, Quantitative Primer and Probe Design, Thermal Cycling Parameters, Sample Preparation, and Real-time Data Analysis. - [Read Real-Time or Kinetic PCR]

Category: Yeast Protocols: Yeast Genetics Protocols: Yeast Replication Protocols

Protocol for replication timing using transient hemimethylation. Includes: Strain construction; Protocol for methylation timing; Quantitation. - [Read Replication Timing Using Transient Hemimethylation Protocol]

Category: Protein Protocols: Protein Quantitation Concentration Protocols

Category: Lipid Protocols

Protocol for sphingomyelin quantitation post-choline labeling of HL60 cells. - [Read Sphingomyelin Quantitation Post-choline Labeling of HL60 Cells Protocol]

Category: Protein Protocols: Protein Quantitation Concentration Protocols

UV Absorbance 280 nm Protein Determination. Simple and quick method to accurately quantitate total protein in purified material or approximately quantitate total protein in crude lysates or partial purified material. Quantitation of the amount of protein in a solution is possible in a simple spectrometer.  Absorption of radiation in the near UV by proteins depends on the Tyr and Trp content (and to a very small extent on the amount of Phe and disulfide bonds).  Dr. Mario Lebendiker - [Read UV Absorbance 280 nm Protein Determination]