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Protocol Immunoprecipitation - http://www.sigmaaldrich.com/Area_of_Interest/Life_Science/Antibody_Explorer/Procedures/Immunoprecipitation.html

Category: Immunology: Immunoprecipitation Protocols

Immunoprecopitation method, the protein from the cell or tissue homogenate is precipitated in an appropriate lysis buffer by means of an immune complex which includes the antigen (protein), primary antibody and Protein A-, G-, or L-agarose conjugate or a secondary antibody-agarose conjugate - [Read Protocol Immunoprecipitation]

Protocol Primary Mammalian Endothelial Cells - http://www.amaxa.com/fileadmin/groups/marketing/Downloads/Protocols/Primary_cells/amaxa_OP_Basic_Endothelial_VPI-1001.pdf

Category: Cell Biology and Cell Culture: Endothelial Cell Protocols

Primary mammalian endothelial cells protocol. This protocol is designed for primary endothelial cells isolated from various organs of mammals. Large and flat cells, often with large nuclei. Includes: Required reagents; DNA preparation and quality; Preparation of cells and cell culture; Important controls; Nucleofection protocol. - [Read Protocol Primary Mammalian Endothelial Cells]

Quantification of Apoptosis and Cell Cycle Distribution of Primary B Cells Using Propidium Iodide - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000024.pdf?pid=PP00000024

Category: Immunology: B Cell Protocols

The water soluble, DNA intercalator, propidium iodide (PI), is used to bind to DNA after permeabilization of cells with NP40. The amount of dye bound correlates with the content of DNA within a given cell. Once cells are stained, they are analyzed on a flow cytometer. The relative content of DNA indicates the distribution of a population of cells throughout the cell cycle. - [Read Quantification of Apoptosis and Cell Cycle Distribution of Primary B Cells Using Propidium Iodide]

Quantification of Apoptosis and the Cell Cycle Distribution of Primary B Cells Using PI - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000024.pdf?pid=PP00000024

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

The water soluble, DNA intercalator, propidium iodide (PI), is used to bind to DNA after permeabilization of cells with NP40. The amount of dye bound correlates with the content of DNA within a given cell. Once cells are stained, they are analyzed on a flow cytometer. The relative content of DNA indicates the distribution of a population of cells throughout the cell cycle. - [Read Quantification of Apoptosis and the Cell Cycle Distribution of Primary B Cells Using PI]

Rat Chromaffin Cells Primary Cultures Protocol - http://www.natureprotocols.com/2006/09/29/rat_chromaffin_cells_primary_c.php

Category: Mouse Protocols: Mouse Cell Culture Protocols

Protocol for rat Chromaffin cells primary cultures: Standardization and quality assessment for single-cell assays. Includes: Preliminars; Isolation of the rat adrenal medulla; Enzymatic digestion of medullary tissue; Collecting and culturing the cells. - [Read Rat Chromaffin Cells Primary Cultures Protocol]

Rat Chromaffin Cells Primary Cultures Standardization and Quality Assessment for Single Cell Assays - http://www.natureprotocols.com/2006/09/29/rat_chromaffin_cells_primary_c.php

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols: Rat Neuronal Cell Culture Protocols

Protocol for standardization of both the isolation procedure and the quality of the cultures. - [Read Rat Chromaffin Cells Primary Cultures Standardization and Quality Assessment for Single Cell Assays]

Rat Chromaffin Cells Primary Cultures: Standardization and Quality Assessment for Single-Cell Assay - http://www.natureprotocols.com/2006/09/29/rat_chromaffin_cells_primary_c.php

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

Protocol to be used as reference to help standardization of both the isolation procedure and the quality of the cultures allowing a better comparison of the results obtained from different laboratories. - [Read Rat Chromaffin Cells Primary Cultures: Standardization and Quality Assessment for Single-Cell Assay]

Rat Neuron Nucleofector Kit - http://www.amaxa.com/fileadmin/groups/marketing/Downloads/Protocols/Primary_cells/amaxa_OP_Neuron_hippo_cortical_rat_DPG-1003.pdf

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols: Rat Neuronal Cell Culture Protocols

Optimized Protocol for primary rat hippocampal or cortical neurons. - [Read Rat Neuron Nucleofector Kit]

Retroviral Infection of Primary Human Keratinocytes (PHKs) Protocol - http://www.methodbook.net/cellcult/infectn.html

Category: Viral Culture and Isolation Protocols: Viral Isolation Protocols

Protocol for the retroviral infection of primary human keratinocytes (PHKs). - [Read Retroviral Infection of Primary Human Keratinocytes (PHKs) Protocol]

Techniques for Primary Culture of C. elegans Embryonic Neurons - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=14

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols: C. elegans Neuronal Cell Culture Protocols

Development of techniques for primary culture of C. elegans embryonic neurons. - [Read Techniques for Primary Culture of C. elegans Embryonic Neurons]

Techniques for Primary Culture of C. elegans Embryonic Neurons II - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=15

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols: C. elegans Neuronal Cell Culture Protocols

Development of techniques for primary culture of C.elegans embryonic neurons. - [Read Techniques for Primary Culture of C. elegans Embryonic Neurons II]

Techniques for Primary Culture of C. elegans Embryonic Neurons III - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=16

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols: C. elegans Neuronal Cell Culture Protocols

Development of techniques for primary culture of C. elegans embryonic neurons - [Read Techniques for Primary Culture of C. elegans Embryonic Neurons III]

Transfecting Cultured Hippocampal Neurons with an Actin-GFP Plasmid - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4664

Category: Transfection Protocols: Calcium Phosphate Transfection Protocols

This protocol describes two transfection methods for expressing GFP-tagged actin in primary neurons. The lipid reagent DOTAP (Roche Diagnostics) method produces actin-GFP-expressing hippocampal neurons that survive well during long periods in culture. The calcium phosphate method can be used to transfect neurons that have already been growing on coverslips in vitro. Transfected cells suitable for imaging can be obtained in cultures up to 15 days in vitro. - [Read Transfecting Cultured Hippocampal Neurons with an Actin-GFP Plasmid]

Transfecting Cultured Hippocampal Neurons with an Actin-GFP Plasmid Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4664

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols

Protocol describes two transfection methods for expressing GFP-tagged actin in primary neurons. The lipid reagent DOTAP (Roche Diagnostics) method produces actin-GFP-expressing hippocampal neurons that survive well during long periods in culture. - [Read Transfecting Cultured Hippocampal Neurons with an Actin-GFP Plasmid Protocol]

Transfection of Bone Marrow-Derived Mast Cells for Transcription Factor Luciferase Reporter Assays - http://www.natureprotocols.com/2006/09/12/transfection_of_bone_marrowder_1.php

Category: Reporter Gene Assays: Luciferase Assay Protocols

Transfection of primary leukocytes has traditionally been a challenging but much desired protocol. It allows not only the analysis of cells in a more natural state to a cell line system, it enables the direct comparison of, for e.g. transcriptional activity using luciferase reporters, in immune cells taken from genetically-altered mice. In addition, importantly it allows for "rescue experiments" in knockout cells & the ability to over-express or reconstitute wild-type and/or mutated constructs. - [Read Transfection of Bone Marrow-Derived Mast Cells for Transcription Factor Luciferase Reporter Assays]

Transfection of Hippocampal Neurons with Plasmid DNA Using Calcium Phosphate Coprecipitation - http://www.cshprotocols.org/cgi/content/abstract/2006/1/pdb.prot4445

Category: Transfection Protocols: Calcium Phosphate Transfection Protocols

This protocol describes transfection of plasmid DNA into primary hippocampal neurons using DNA/calcium-phosphate (CaPO4) coprecipitation. The precise pH of the transfection medium and the incubation time of cells with the coprecipitate are critical for reproducible and efficient transfection. Once these parameters are optimized for a given plasmid, the method is easily adapted for transfection of other established cell lines. - [Read Transfection of Hippocampal Neurons with Plasmid DNA Using Calcium Phosphate Coprecipitation]

VEGF Antibody Staining Protocol - http://www.ihcworld.com/_protocols/antibody_protocols/vegf_vg1_chemicon.htm

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

Protocol for VEGF antibody staining. Includes: Primary Antibody; Antigen Retrieval; Detection Methods; Chromogen Substrate; Counterstain. - [Read VEGF Antibody Staining Protocol]

Worthington Tissue Dissociation Guide - http://www.tissuedissociation.com/techniques.html

Category: Cell Culture Protocols

Includes information on: Cell Isolation Theory ( Tissue Types: Epithelial Tissue, Connective Tissue); Dissociating Enzymes (Collagenase, Trypsin, Elastase, Hyaluronidase, Papain, Chymotrypsin, Deoxyribonuclease I, Neutral Protease (Dispase), Trypsin Inhibitor (Soybean)); Cell Isolation Techniques: Working With Enzymes, Basic Primary Cell Isolation, Equilibration with 95%O2:5%CO2, Trituration, Enzymatic Cell Harvesting, Cell Adhesion and Harvesting, Trypsin for Cell Harvesting, etc... - [Read Worthington Tissue Dissociation Guide]

Articles (1-5 of 5)

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

Tubulin Polymerization Protocol using GTP and GMPCPP

Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.

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