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Annexin V Protocol - http://www.cyto.purdue.edu/flowcyt/research/cytotech/apopto/data/chap16.htm

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

Annexin V, belonging to a recently discovered family of proteins, the annexins, with anticoagulant properties has proven to be a useful tool in detecting apoptotic cells since it preferentially binds to negatively charged phospholipids like PS in the presence of Ca2+ and shows minimal binding to phosphatidylcholine and sphingomyeline. Changes in PS asymmetry, which is analyzed by measuring Annexin V binding to the cell membrane, were detected before morphological changes associated with... - [Read Annexin V Protocol]

Bradford Assay Method - http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Biochemistry/Bradford.html

Category: Protein Protocols: Protein Quantitation Concentration Protocols: Bradford Protein Assay Protocols

The bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie Brilliant blue to protein. There is no interference from cations nor from carbohydrates such as sucrose.
Detergents such as sodium dodecyl sulfate and triton x-100 can interfere with the assay, as well as strongly alkaline solutions.

Includes a general overview of the procedure and preparation of the standards in the protocol. - [Read Bradford Assay Method]

Drosophila Larval Chemotaxis Assays - http://vosshall.rockefeller.edu/protocols/LarvalChemotaxisAssay.pdf

Category: Immunology: Chemotaxis

Drosophila Larval Chemotaxis Assays. Larva harvesting. Olfactory tests on control larvae. Measuring response. Leslie Vosshall Laboratory, Rockefeller University, New York. - [Read Drosophila Larval Chemotaxis Assays]

Early Events in B Lymphocyte Activation Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E6634190D367003ECF94AE65295FA77&objectid=6674AB8295693316922D8DFC029193A6

Category: Immunology: B Cell Protocols

B cell activation can be quantitated indirectly by assaying antibody production or directly by measuring cellular changes that occur immediately after exposure to an activation signal. Provides methods for the latter (direct) approach--namely, methods for quantifying early parameters of B cell activation such as increases in intracellular ionized calcium concentration [Ca2+]I, cell size, and MHC class II-antigen expression. - [Read Early Events in B Lymphocyte Activation Protocol]

ELISA Method Measure Inhibition COX Enzymes - http://www.natureprotocols.com/2006/11/22/an_elisa_method_to_measure_inh.php

Category: Pharmacology and Toxicology Protocols: Enzyme Assays Protocols

Protocol describes a procedure measuring cyclooxygenase activity by quantifying PGE2 produced by enzymatic conversion of arachidonic acid, in the presence or absence of potential inhibitors. This high-throughput method has the advantage that it directly measures cyclooxygenase activity and requires little enzyme. - [Read ELISA Method Measure Inhibition COX Enzymes]

ELISA Method to Measure Inhibition of the COX Enzymes Protocol - http://www.natureprotocols.com/2006/11/22/an_elisa_method_to_measure_inh.php

Category: ELISA Protocols

The cyclooxygenase (COX) reaction can be monitored by measurement of oxygen consumption, peroxidase co-substrate oxidation or prostaglandin (PG) detection. This protocol describes a procedure measuring cyclooxygenase activity by quantifying PGE2 produced by enzymatic conversion of arachidonic acid, in the presence or absence of potential inhibitors. - [Read ELISA Method to Measure Inhibition of the COX Enzymes Protocol]

ELISA Procedure for Measuring Serum Antibody Titer - http://www.komabiotech.com/technical/protocol/ELISA.htm

Category: ELISA Protocols

Method and ELISA Procedure for Measuring Serum Antibody Titer. Komabiotech - [Read ELISA Procedure for Measuring Serum Antibody Titer]

Imaging and Measuring Biomolecules & Their Assemblies by Scanning Transmission Electron Microscopy - http://www.mih.unibas.ch/Booklet/Booklet96/Chapter4/Chapter4.html

Category: Microscopy Protocols: Electron Microscopy Protocols

The scanning transmission electron microscope precision and reproducibility of mass measurements are comparable with those of the analytical ultracentrifuge, the possibility of determining the mass not only of entire supramolecular assemblies but also of their distinct components has opened exciting new avenues which have occasionally been entered but are not yet fully explored. Includes: Principle and application (The GroEL:GroES complex). - [Read Imaging and Measuring Biomolecules & Their Assemblies by Scanning Transmission Electron Microscopy]

Imaging, Measuring and Manipulating Native Biomolecular Systems with the Atomic Force Microscope - http://www.mih.unibas.ch/Booklet/Booklet96/Chapter3/Chapter3.html

Category: Microscopy Protocols

The atomic force microscope (AFM) is one of the most powerful tools for determining the surface topography of native biomolecules at subnanometer resolution. The AFM can also provide insight into the binding properties of biological systems. In order to determine the specific interaction between two kinds of molecules (e.g., avidin and biotin). Includes information on principle of AFM and application of AFM. - [Read Imaging, Measuring and Manipulating Native Biomolecular Systems with the Atomic Force Microscope]

Immunohistochemistry using Anti-Ganglioside Antibodies - http://www.glycotech.com/protocols/Proto2.html

Category: Molecular Biology Protocols: Carbohydrate Protocols

Indirect method measuring immunofluorescence coupled to second antibody. Best for membrane antigens in addition to intra- and extracellular antigens, may be applied to frozen tissue sections, to cells in suspension, and to cells attached to glass slides or coverslips. Tadashi Tai~Head, Department of Tumor Immunology, The Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan - [Read Immunohistochemistry using Anti-Ganglioside Antibodies]

Measurement of GFP Expression and DNA Content in Permeabilized Cells - http://cyto.mednet.ucla.edu/Protocols/DNA%20and%20GFP%20PI.htm

Category: Reporter Gene Assays: GFP Assay Protocols

Protocol measuring the amount of GFP expression and DNA content in permeabilized cells. Includes: Fix cells with formaldehyde; Permeabilize cells with ethanol; Preparation of Buffered 2% Formaldehyde Solution. - [Read Measurement of GFP Expression and DNA Content in Permeabilized Cells]

Measuring Apoptosis and Necrosis by Dual Laser Flow Cytometry Protocol - http://cyto.mednet.ucla.edu/Protocols/simultta.htm

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]

Measuring Apoptosis and Necrosis by Dual Layer Flow Cytometry - http://cyto.mednet.ucla.edu/Protocols/simultta.htm

Category: Cell Biology and Cell Culture: Apoptosis Protocols

Measuring Apoptosis and Necrosis (Oncosis) by Dual Layer Flow Cytometry - [Read Measuring Apoptosis and Necrosis by Dual Layer Flow Cytometry]

Measuring Protein Concentration by Western Analysis Using Enhanced Chemiluminescence Detection - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4255

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]

Measuring Volume Protocol - http://homepages.gac.edu/~cellab/chpts/chpt1/ex1-6.html

Category: Microscopy Protocols

Protocol for measuring volume. Material required includes: Microscope, Hemacytometer and coverslip, Suspension of yeast. - [Read Measuring Volume Protocol]

Measuring Yeast Cell Density by Spectrophotometry Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4186

Category: Yeast Protocols

Simple method for determining the cell density of yeast cultures. - [Read Measuring Yeast Cell Density by Spectrophotometry Protocol]

Mechanical Tracing of Protein Function in the Drosophila Ear Protocol - http://www.natureprotocols.com/2006/11/03/mechanical_tracing_of_protein.php

Category: Drosophila Protocols

Method relies on the examination of the ear’s mechanics, which is actively modulated by the motility of auditory neurons and reflects the function of mechanosensory proteins these cells comprise [5-7]. Mechanical signatures arising from the motility of the neurons are assayed by measuring the vibrations of the antennal sound receiver in the presence and absence of sound. - [Read Mechanical Tracing of Protein Function in the Drosophila Ear Protocol]

NIH Image for Mac - http://rsb.info.nih.gov/nih-image/

Category: Molecular Imaging

NIH Image. Image can be used to measure area, mean, centroid, perimeter, etc. of user defined regions of interest. It also performs automated particle analysis and provides tools for measuring path lengths and angles. Spatial calibration is supported to p - [Read NIH Image for Mac]

Photoactivation-Based Labeling and In Vivo Tracking of RNA Molecules in the Nucleus - http://www.cshprotocols.org/cgi/content/abstract/2006/29/pdb.prot4600

Category: Microscopy Protocols: Live-Cell Imaging Protocols

This protocol describes a method for observing and measuring the movement of RNA molecules in the nucleus of living mammalian cells. Caged fluorescein-labeled DNA oligonucleotides are introduced into living mammalian cells, where they demonstrably hybridize to complementary RNA. After site-specific photoactivation at desired sites within the cell, the RNA movements away from those sites are followed and digitally recorded using a rapid acquisition microscopy system. - [Read Photoactivation-Based Labeling and In Vivo Tracking of RNA Molecules in the Nucleus]

Photosynthesis and Respiration - Introduction - http://homepages.gac.edu/~cellab/chpts/chpt8/intro8.html

Category: Plant Biology Protocols: Photosynthesis and Respiration Protocols

The physiological reactions of mitochondria and chloroplasts can be reduced to a series of electron transfers, catalyzed by specific enzymes found within the organelles. Thus, we can study the component processes of photosynthesis and respiration by isolating the organelles and measuring specific enzyme activity associated with that organelle. - [Read Photosynthesis and Respiration - Introduction]

Preparation of B-Lymphocyte Lysates for Cyclic AMP Determination - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000012.pdf?pid=PP00000012

Category: Signalling Signal Transduction Protocols

This protocol provides a sufficient sample for several determinations of cAMP using the acetylation protocol. The method chosen for measuring the content of cyclic adenosine 3',5'-monophosphate (cyclic AMP or cAMP) in splenic B lymphocytes (B cells) is an enzyme linked immunoassay system. Protocol includes information on: Treatment of Cells and Preparation of Extracts; Reagents and Materials. - [Read Preparation of B-Lymphocyte Lysates for Cyclic AMP Determination]

Preparation of B-Lymphocyte Lysates for Cyclic AMP Determination for Dual Ligand Screen Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000139.pdf?pid=PP00000139

Category: Immunology: B Cell Protocols

The method chosen for measuring the content of cyclic adenosine 3',5'- monophosphate (cyclic AMP or cAMP) in splenic B lymphocytes (B cells) is an enzymelinked immunoassay system. Includes: Treatment of Cells and Preparation of Extracts. - [Read Preparation of B-Lymphocyte Lysates for Cyclic AMP Determination for Dual Ligand Screen Protocol]

Preparation of Myocyte Lysates for Cyclic AMP Determination - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000131.pdf?pid=PP00000131

Category: Signalling Signal Transduction Protocols

Protocol provides a method for acheiving a sufficient sample for several determinations of cAMP. The protocol described for measuring the content of cyclic adenosine 3',5'- monophosphate (cyclic AMP or cAMP) in cardiac myocytes is an enzyme-linked immunoassay system. Protocol includes information on: Treatment of Cells and Preparation of Extracts; Use of Environmental Chamber; Reagents and Materials. - [Read Preparation of Myocyte Lysates for Cyclic AMP Determination]

Preparation of RAW 264.7 Lysates for Cyclic AMP Determination - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000175.pdf?pid=PP00000175

Category: Signalling Signal Transduction Protocols

The protocol provides a method to achieve a sample sufficient for one determination of cAMP using the acetylation protocol. The protocol describes the method used for measuring the content of cyclic adenosine 3',5'- monophosphate (cyclic AMP or cAMP) in RAW 264.7 cells using an enzyme-linked immunoassay system. Information included in the protocol: Treatment of Cells and Preparation of Extracts; Reagents and Materials. - [Read Preparation of RAW 264.7 Lysates for Cyclic AMP Determination]

Proliferative Assays for B Cell Function Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E663429ABD843E3419A0C3061FDD3E4&objectid=6674AEBEC084AF4DB9D3826E3ED9B9A5

Category: Immunology: B Cell Protocols

Describes procedures for measuring the capacity of purified B cells to undergo proliferation. The method centers on the use of polyclonal stimulating agents (mitogens) because these agents stimulate the majority of B cells and because the alternative (measurement of antigen-induced proliferation) requires the laborious procedures of isolating antigen-specific B cells (which are otherwise present in too low a concentration in whole B cell populations). - [Read Proliferative Assays for B Cell Function Protocol]

Articles (1-4 of 4)

Genomic DNA Labeling of Microarrays Protocol

DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.

Acid Guanidinium Thiocyanate RNA Isolation Phenol Chloroform Extraction

A single step RNA isolation protocol using Phenol Chloroform Extraction and Acid Guanidinium Thiocyanate. This RNA isolation method uses the fact that guanidinium thiocyanate can simultaneously lyse the cells and inactive cellular RNAses during the initial RNA isolation step allow a single step in the method.