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DNA Affinity Chromatography Using Gravity Flow - Subscription Required - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4206

Category: Chromatography Protocols: DNA RNA Affinity Chromatography

DNA Affinity Chromatography, DNA affinity chromatography can be a low-tech method using gravity flow at 4Â°C, a disposable chromatography column, and DNA affinity resin prepared in the laboratory (see Preparation of a DNA Affinity Column). Include 10-20% glycerol and 0.025-0.1% NP-40 in the column buffers to suppress losses due to nonspecific adsorption of protein to surfaces. Load the protein in a buffer that is compatible with binding of the protein to its target site. Keith Brocklehurst et al - [Read DNA Affinity Chromatography Using Gravity Flow - Subscription Required]

DNA Analysis by Flow Cytometry Protocol - http://www.niehs.nih.gov/research/atniehs/core/fc/protocols/dna.cfm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol for DNA analysis by flow cytometry. - [Read DNA Analysis by Flow Cytometry Protocol]

DNA laddering assay for treated cells - http://www.celldeath.de/apometh/dnaladd.html

Category: Cell Biology and Cell Culture: Apoptosis Protocols

DNA laddering can be detected from samples with only 8% apoptotic cells. Alternatively, the cells can be stained with DAPI and analysed by flow cytometry. CellDeath.de - [Read DNA laddering assay for treated cells]

Dynamic Flow Assay for Cell Adhesion in a Parallel Plate Flow Chamber - http://www.glycotech.com/protocols/proto7.html

Category: Cell Biology and Cell Culture

Flow assays offer visualization of cell adhesion under wall shear stress. Visualization of the different events of cell adhesion can be quantified by selective image acquisition and subsequent image processing. Flow assays are suited for adhesive events which occur very rapidly in a time scale shorter than that of most static adhesion assays. Also, events subsequent to the initial events can be studied such as cell stabilization and spreading giving some insight into the kinetics of cell-cell. - [Read Dynamic Flow Assay for Cell Adhesion in a Parallel Plate Flow Chamber]

Dynamic Flow Assay in a Parallel Plate Flow Chamber - http://www.glycotech.com/protocols/proto7.html

Category: Cell Biology and Cell Culture: Cell Adhesion Protocols

Flow assays allow visualization of cell adhesion under well-defined wall shear stress. Visualization of the events of cell adhesion are quantified by selective image acquisition and image processing. Events subsequent to the initial events can be studied such as cell stabilization and spreading. John T. Patton~GlycoTech Corporation, Rockville, Maryland - [Read Dynamic Flow Assay in a Parallel Plate Flow Chamber]

Estimating the Number of CD38 Molecules on the CD8+ T Lymphocytes of HIV-Infected Individuals - http://cyto.mednet.ucla.edu/Protocols/cd38.pdf

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]

FIXATION PROTOCOL FOR FLOW CYTOMETRIC ANALYSIS OF DNA BY PI STAINING - http://www.vetmed.auburn.edu/index.pl/cell_cycle/dna_content

Category: Histology Protocols: Fixation of Cells Tissues Protocol

Analysis of Cell Cycle Based on DNA Content. FIXATION PROTOCOL FOR FLOW CYTOMETRIC ... Liberate cells from tissue culture flask by trypsin digestion and ... FIXATION PROTOCOL FOR FLOW CYTOMETRIC ANALYSIS OF DNA BY PI STAINING. Auburn University, College of Veterinary Medicine. You. - [Read FIXATION PROTOCOL FOR FLOW CYTOMETRIC ANALYSIS OF DNA BY PI STAINING]

Flow Cell Assays with Microtubules: Motility/Dynamics in Fluorescence and VE-DIC Protocol - http://mitchison.med.harvard.edu/protocols/flowcell.html

Category: Cytoskeleton Protocols: Microtubules Protocols

Protocol for flow cell assays with microtubules. Includes: Solutions & Supplies; VE-DIC Assays; Motility; Dynamics; Fluorescence Assays; Oxygen Scavenging; Assays. - [Read Flow Cell Assays with Microtubules: Motility/Dynamics in Fluorescence and VE-DIC Protocol]

Flow Cytometric Analysis of Gram-Negative Bacterial Cells - http://www.cyto.purdue.edu/flowcyt/research/micrflow/robert/robert.htm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Flow cytometric analysis of gram-negative bacterial cells. Includes: Staining Conditions and FACSort Settings; Light Scatter Resolution on the BD FACSort; Bacterial Cell Resolution on the BD FACSort. - [Read Flow Cytometric Analysis of Gram-Negative Bacterial Cells]

Flow Cytometric Analysis of Mitochondrial Transmembrane Potential ({Delta}{Psi}m) - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4465

Category: Cell Organelle Protocols: Mitochondria Protocols

Protocol describes a method for evaluation of mitochondrial function using the fluorochrome CMXRos. CMXRos is sequestered by actively respiring mitochondria, but washed out when the mitochondrial membrane potential is lost. This analysis can be combined with the TUNEL technique or immunocytochemistry. - [Read Flow Cytometric Analysis of Mitochondrial Transmembrane Potential ({Delta}{Psi}m)]

Flow cytometric determination of leukocyte surface antigens in whole blood - http://pingu.salk.edu/flow/protocols/wholeblood.html

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]

Flow Cytometric Determination of Leukocyte Surface Antigens in Whole Blood Protocol - http://pingu.salk.edu/flow/protocols/wholeblood.html

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]

Flow Cytometry Analysis Protocol - http://www.cbrinstitute.org/labs/springer/protocols/qing_facs.html

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

Flow cytometry is a widely used method for characterizing and separating individual cells. This basic protocol focuses on: measure fluorescence intensity produced by fluorescent-labled antibodies and ligands that bind specific cell-associated molecules. Includes: Immunofluorescence Staining and Flow Cytometry Analysis. - [Read Flow Cytometry Analysis Protocol]

Flow Cytometry Analysis Protocol - http://www.cbrinstitute.org/labs/springer/protocols/qing_facs.html

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Flow cytometry is a widely used method for characterizing and separating individual cells. This basic protocol focuses on: measure fluorescence intensity produced by fluorescent-labled antibodies and ligands that bind specific cell-associated molecules. Includes: Immunofluorescence Staining; Flow Cytometry Analysis. - [Read Flow Cytometry Analysis Protocol]

Flow Cytometry Analysis Protocol - http://cbr.med.harvard.edu/labs/springer/protocols/qing_facs.html

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

Flow Cytometry Analysis Protocol. Springer Lab - Harvard. Background on flow cytometry technique: Flow cytometry is a method which uses instrumentation scanning single cells flowing past excitation sources in a liquid medium.
- [Read Flow Cytometry Analysis Protocol]

Flow Cytometry Laboratory - http://www.cyto.purdue.edu/cdroms/flow/vol3/8/data/icrf/apop.htm

Category: Cell Biology and Cell Culture: Apoptosis Protocols

Flow Cytometry Laboratory. Apoptosis. Perdue. - [Read Flow Cytometry Laboratory]

Flow Cytometry of Apoptosis Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E663022BBACCC599F244293812ED95A&objectid=66744E3FA79C848C2D8B85BB84708AC9

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

Common methods applicable to flow cytometry make it possible to: (1) identify and quantify dead or dying cells, (2) reveal a mode of cell death (apoptosis or necrosis), and (3) study mechanisms involved in cell death. Gross changes in cell morphology and chromatin condensation, which occur during apoptosis, can be detected by analysis with laser light beam scattering. - [Read Flow Cytometry of Apoptosis Protocol]

Fluorescein Labeling of Proteins Protocol - http://pingu.salk.edu/flow/protocols/flabel.html

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

Protocol for fluorescein labeling of proteins. - [Read Fluorescein Labeling of Proteins Protocol]

Fluorescent Dyes for Flow Cytometric Analysis - http://www.ebioscience.com/ebioscience/appls/Dyes.htm

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

Fluorescent dyes absorb light at certain wavelengths and in turn emit their fluorescence energy at a higher wavelength. Each dye has a distinct emission spectrum, which can be exploited for multicolor analysis. eBioscience antibodies are available conjugated to a wide variety of fluorochromes. - [Read Fluorescent Dyes for Flow Cytometric Analysis]

Fluorescent Staining of Cell Surface Antigens on Viable Cells Using Direct Immunofluorescence - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4439

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

Protocol provides a method for analysis of cell surface antigens using direct immunofluorescence, i.e., when the antibody being used is directly labeled to a fluorescent dye. After staining, the cells can be analyzed immediately by flow cytometry or fixed and stored for later analysis. - [Read Fluorescent Staining of Cell Surface Antigens on Viable Cells Using Direct Immunofluorescence]

Gene Delivery by Direct Injection (Microinjection) Using a Controlled-Flow System Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4654

Category: Microinjection Protocols: Microinjection of DNA Protocols

This protocol describes a method for constant-flow microinjection using the Pneumatic PicoPump (World Precision Instruments). This type of system is very simple and can be assembled on a relatively low budget. In this method, a constant flow of sample is delivered from the tip of the pipette, and the amount of sample injected into the cell is determined by how long the pipette remains in the cell. - [Read Gene Delivery by Direct Injection (Microinjection) Using a Controlled-Flow System Protocol]

Gene Delivery by Direct Injection (Microinjection) Using a Pulsed-Flow System Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4653

Category: Microinjection Protocols: Microinjection of DNA Protocols

This protocol describes a method for pulsed-flow microinjection using the Eppendorf FemtoJet injector and Eppendorf InjectMan; this is the most common type of pulsed-flow microinjection system currently being used. The advantage of this type of system over a controlled-flow system is that much more control is available over the injection parameters, reducing variability in injections. In addition, the system allows a diagonal insertion of the needle into the cell. - [Read Gene Delivery by Direct Injection (Microinjection) Using a Pulsed-Flow System Protocol]

Immunodetection of cyclin D1 and D2/D3 using 488/630 nm dual laser flow cytometry Protocol - http://www.cyto.purdue.edu/flowcyt/research/cytotech/telford/cyclinip.htm

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

Protocol for immunodetection of cyclin D1 and D2/D3 using 488/630 nm dual laser flow cytometry. - [Read Immunodetection of cyclin D1 and D2/D3 using 488/630 nm dual laser flow cytometry Protocol]

Immunofluorescent Staining for Flow Cytometry Protocols - http://www.ebioscience.com/ebioscience/appls/FCS.htm

Category: Immunology: Immunofluorescence Protocols: Immunofluorescence Staining Protocols

Protocols for immunofluorescent staining for flow cytometry. Includes: Protocol for Staining of Cell Suspensions of Murine Lymphoid Tissue; Protocol for Staining of Human Peripheral Blood; Fluorescent Dyes for Flow Cytometric Analysis - [Read Immunofluorescent Staining for Flow Cytometry Protocols]

Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis - http://www.bdbiosciences.com/pharmingen/protocols/Intracellular_Cytokines.shtml

Category: Signalling Signal Transduction Protocols: Cytokine Protocols

Fluorescent anti-cytokine and monoclonal antibodies are useful for the intracellular staining and multiparameter flow cytometric analysis of individual cytokine-producing cells within mixed cell populations. BD Biosciences. - [Read Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis]

Articles (1-3 of 3)

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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