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5-Bromo-2-Deoxyuridine (BrdU) and 7-Amino-Actinomycin (7-AAD) Staining for Cell Proliferation Assay - http://www.natureprotocols.com/2007/01/22/5bromo2deoxyuridine_brdu_and_7.php

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: DNA Staining Protocols for Flow Cytometry

The incorporation of 5-bromo-2-deoxyuridine (BrdU) in replicating DNA of proliferating cells can be used to measure their proliferation. This protocol allows the identification of cells which have incorporated BrdU by flow cytometry. - [Read 5-Bromo-2-Deoxyuridine (BrdU) and 7-Amino-Actinomycin (7-AAD) Staining for Cell Proliferation Assay]

A sensitive method for detection of apoptosis by single laser FACS - http://cyto.mednet.ucla.edu/Protocols/detection.htm

Category: Cell Biology and Cell Culture: Apoptosis Protocols

A sensitive method for detection of apoptosis by single laser Flow Cytometry. - [Read A sensitive method for detection of apoptosis by single laser FACS]

Advanced Methodologies in Flow Cytometry - http://www.cyto.purdue.edu/flowcyt/research/cytotech/amfc/data/index1.htm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Data Analysis Protocols

Advanced Methodologies in Flow Cytometry. Andrea Cossarizza et al.. Materials from a workshop held March, 1997 in Modena, Italy. - [Read Advanced Methodologies in Flow Cytometry]

Amplification of Flow Sort DNA using DOP-PCR (SKY) Protocol - http://www.riedlab.nci.nih.gov/publications/AmpFlowSortDNA_DOP-PCR.pdf

Category: Cytogenetics Protocols: Spectral Karyotyping SKY Protocols

Protocol for the amplification of flow sort DNA using DOP-PCR (SKY). - [Read Amplification of Flow Sort DNA using DOP-PCR (SKY) Protocol]

Analysis of Cellular DNA Content by Flow Cytometry Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E661C8A9C23374B5FAB0610666CCDFB&objectid=6674F58BDEF6DCA5276258FEA4CD534D

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: DNA Staining Protocols for Flow Cytometry

Simple and universally applicable methods for staining fixed cells are presented, as are methods that utilize detergents and proteolytic treatment to permeabilize cells. Additionally, supravital cell staining with Hoechst 33342, which is primarily used for sorting live cells for subsequent culturing based on DNA-content differences, is also described. Also presented are methods for staining of cell nuclei isolated from paraffin-embedded tissues, and deconvolution of DNA-content-frequency... - [Read Analysis of Cellular DNA Content by Flow Cytometry Protocol]

Analysis of DNA Fragmentation Using Propidium Iodide (PI) Fluorescence of Individual Nuclei Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4430

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: DNA Staining Protocols for Flow Cytometry

This protocol provides a method for analysis of DNA fragmentation using flow cytometry after propidium iodide (PI) labeling of apoptotic nuclei. Apoptotic nuclei stain less than their normal counterparts because of diffusion of low-molecular-weight fragments out of the cells and/or chromatin condensation. - [Read Analysis of DNA Fragmentation Using Propidium Iodide (PI) Fluorescence of Individual Nuclei Protocol]

Analysis of DNA Fragmentation Using Propidium Iodide (PI) Staining After Ethanol Fixation Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4431

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: DNA Staining Protocols for Flow Cytometry

This protocol provides a method for analysis of DNA fragmentation using flow cytometry after propidium iodide (PI) labeling of fixed apoptotic cells. Flow cytometry reveals apoptotic nuclei in the subdiploid region of the cell cycle histogram... - [Read Analysis of DNA Fragmentation Using Propidium Iodide (PI) Staining After Ethanol Fixation Protocol]

Analysis of Flow Cytometry Data Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E661C2CE22FCCB1C294CD8376FD8830&objectid=6674E762AC837B13929440A1F32AAEF0

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Data Analysis Protocols

Provides several approaches to flow cytometry data analysis. Frequency determinations based on analysis of single-parameter fluorescence histograms and dual-parameter contour plots are presented. Steps are described for calculating values for signal-to-noise ratios when logarithmic amplification is used for data collection. - [Read Analysis of Flow Cytometry Data Protocol]

Annexin V Protocol for Flow Cytometry - http://www.niehs.nih.gov/research/atniehs/core/fc/protocols/annexin.cfm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Different cell types vary in their phosphatidylserine (PS) content, along with the amount of PS exposure on the cell surface after cell death. This protocol is a guideline for getting started, however it may be necessary to adjust the concentration of the Annexin V-FITC. - [Read Annexin V Protocol for Flow Cytometry]

Annexin V Staining Flow Cytometry Protocol - http://www.cyto.purdue.edu/flowcyt/research/cytotech/amfc/data/page1.htm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Annexin V Staining Flow Cytometry Protocol - a general protocol for staining cell for cytometry analysis. BD PharMingen. - [Read Annexin V Staining Flow Cytometry Protocol]

Apoptosis Detection Protocol By Single Laser Flow Cytometry - http://cyto.mednet.ucla.edu/Protocols/detection.htm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

A sensitive method for the detection of apoptosis by single laser flow cytometry. Methodology includes: Staining for detection of apoptosis, Direct Staining Procedure, Indirect Staining Procedure, Protocol for the use of actinomycin D (AD) on samples that were stained with 7-AAD for apoptosis and fixed in formaldehyde. - [Read Apoptosis Detection Protocol By Single Laser Flow Cytometry]

Basic Method for Direct Immunofluorescence Labeling - http://www.biotech.iastate.edu/facilities/CELLHYB/Labeling.htm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Basic Method for Direct Immunofluorescence Labeling. Iowa State University, Flow Cytometry Facility. - [Read Basic Method for Direct Immunofluorescence Labeling]

Calcium Flux Analysis Protocol for Flow Cytometry - http://www.niehs.nih.gov/research/atniehs/core/fc/protocols/calcium.cfm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol for calcium flux analysis. Includes: Reagent Preparation; Sample Preparation; Cell Surface Staining; Instrument Set up; Optimizing the Instrument Settings; Recording Data to Apply Compensation; Recording Experimental Data. - [Read Calcium Flux Analysis Protocol for Flow Cytometry]

Calcium Flux Analysis Protocol for Flow Cytometry - http://www.niehs.nih.gov/research/atniehs/core/fc/protocols/calcium.cfm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Intracellular Ion Measurement Protocols

Protocol for calcium flux analysis for flow cytometry. - [Read Calcium Flux Analysis Protocol for Flow Cytometry]

Calibration of Becton Dickinson Flow Cytometers for Relative Fluorescence Intensity Measurements - http://cyto.mednet.ucla.edu/Protocols/relative.htm

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

Flow cytometers must be calibrated prior to fluorescence intensity measurements because of inherent instrument variability. To correct for this variability, a standard particle (fixed chicken red blood cells, or CRBCs) must be analyzed on the instrument prior to each experiment and photomultiplier tube (PMT) voltages adjusted accordingly to place the CRBC fluorescence emission peaks into predetermined target channels. - [Read Calibration of Becton Dickinson Flow Cytometers for Relative Fluorescence Intensity Measurements]

CaspaTag Caspase Activity Protocol - http://www.niehs.nih.gov/research/atniehs/core/fc/protocols/caspatag.cfm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

This assay is based on carboxyfluorescein labeled fluoromethyl ketone (FMK)-peptide inhibitors of caspases. Includes: Working Dilution of FMK-peptide inhibitors; 1X Working Dilution Wash Buffer; Protocol for Flow Cytometry. - [Read CaspaTag Caspase Activity Protocol]

Cell Cycle Protocol using Flow Cytometry - http://www.people.cornell.edu/pages/jls269/protocols/cellcycleprotocols.html

Category: Cell Biology and Cell Culture: Cell Cycle Protocols

Cell Cycle Protocol using Flow Cytometry. Cell cycle Analysis by Flow Cytometry. DNA content Analysis on the FACSAria. Analyzing Cell cycle FCM data. Biomedical Sciences Flow Cytometry Core Laboratory. Dr.Smith Cornell. - [Read Cell Cycle Protocol using Flow Cytometry]

Cell Surface Labeling Protocol - http://sciencepark.mdanderson.org/fcores/flow/files/Cell%20Surface%20Labeling.pdf

Category: Fluorescent Dyes Protocols

Protocol for cell surface labeling. - [Read Cell Surface Labeling Protocol]

Characterization of Cells by Flow Cytometry Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000018.pdf?pid=PP00000018

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol uses specific antibodies coupled to one of four fluorochromes: fluorescein isothiocyanate (FITC), R-phycoerythrin (PE), peridinin chlorophyll-a (PcP), and allophycocyanin (APC). These fluorochromes can be used simultaneously to stain and analyze the expression patterns of four different proteins in the same sample. The fluorochrome stained cell populations are analyzed using a FACSCalibur dual-laser flow cytometer. - [Read Characterization of Cells by Flow Cytometry Protocol]

Characterization of Cells by Flow Cytometry Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000018.pdf?pid=PP00000018

Category: Immunology: B Cell Protocols

Fluorochromes can be used simultaneously to stain and analyze the expression patterns of four different proteins in the same sample. The fluorochrome stained cell populations are analyzed using a FACSCalibur dual-laser flow cytometer. - [Read Characterization of Cells by Flow Cytometry Protocol]

Chelating Sepharose Fast Flow Protocol - http://www.bio.indiana.edu/~ybelab/procedures/ChelatingSepharose.pdf

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Immobilized metal ion affinity chromatography (IMAC) exploits a molecule’s affinity for chelated metal ions. The amino acid histidine present in many proteins forms complexes with transition metal ions such as Cu2+, Zn2+, Ni2+ and Fe3+. Chelating Sepharose™ Fast Flow with a suitable immobilized metal ion will therefore selectively retain proteins with exposed histidine. - [Read Chelating Sepharose Fast Flow Protocol]

Chemokine Receptor Expression Assay by Flow Cytometry - http://ubik.microbiol.washington.edu/protocols/facs/chemokinereceptorfacs.htm

Category: Signalling Signal Transduction Protocols: Chemokine Protocols

Chemokine Receptor Expression Assay by Flow Cytometry. Mullins Lab. Reagents, Protocol and detailed procedure. - [Read Chemokine Receptor Expression Assay by Flow Cytometry]

Configuration Column Construction Column Packing for Capillary Liquid Chromatography - http://cshprotocols.cshlp.org/cgi/content/abstract/2007/3/pdb.prot4578

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol for configuration, column construction, and column packing for a capillary liquid chromatography system. Protocol describes a procedure for adapting conventional HPLC systems to provide accurate low-flow rates (0.4-4 µl/min) and gradients required to operate slurry-packed capillary columns. A key component of this system is a commercial axial-beam longitudinal flow cell that can be fitted to several commercial UV detectors. - [Read Configuration Column Construction Column Packing for Capillary Liquid Chromatography]

Confocal Microscopy - Simple Definitions Queens Univ. - http://meds.queensu.ca/qcri/flow/imageanalysis.htm

Category: Molecular Imaging: Confocal Microscopy Protocols and Information

Confocal Microscopy - Simple Definitions Queens Univ.Image Analysis, Co-localization Analysis, 3D Reconstruction, Object Tracking, Timelapse - Confocal, Deconvolution. - [Read Confocal Microscopy - Simple Definitions Queens Univ.]

Determining Cell Cycle Stages by Flow Cytometry Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E662F660A11AB3ABD97E38B90B2AF0D&objectid=6674031CC5AE0F442766156EA393FE81

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Cycle Analysis Protocols

Describes assays used to determine the distribution of a population of cells to the different stages of the cell cycle as analyzed by flow cytometry. Staining the DNA with different fluorescent dyes, propidium iodide or DAPI, is one of the most direct ways of staging the cells based on DNA content. - [Read Determining Cell Cycle Stages by Flow Cytometry Protocol]

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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.

Coupling Cysteine Containing Peptides to Carrier Protein for Immunization and Polyclonal Antibody

This protocol is used in the production of polyclonal antibodies that recognize a peptide sequence of interest.

Picking Transfected ES Cell Clones Protocol

This protocol a protocol on how to generate transfected embryonic stem (ES) cell clones. The previous protocol in this series is the Protocol for Electroporation of ES cells. The next protocol in the series is the Protocol on Disaggregation, Expansion, and Freezing of Transfected ES Clones.

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