support Protocols

Category: Microscopy Protocols: Live-Cell Imaging Protocols

Protocol on the details required of support slide construction for a sealed preparation for long-term observations of cultured cells. - [Read A Sealed Preparation for Long-Term Observations of Cultured Cells: Support Slide Construction]

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols

Basic Ideas in PCR Protocol Design. PDF - [Read Basic Ideas in PCR Protocol Design. PDF]

Category: PCR Protocols

Basic Ideas in PCR Protocol Design. PDF - [Read Basic Ideas in PCR Protocol Design. PDF]

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

ChIP Staph A cell preparation, preblocking and ChIP IP preclear, recovery, wash steps based on Farnham Lab Protocols. - [Read Chromatin Immunoprecipitation Protocol]

Category: Epigenetics and Methylation Protocols: ChIP Chromatin Immunoprecipitation Protocols

Chromatin Immunoprecipitation Protocol for Microarray Analysis – Protein A/G Bead Method. UHN Microarray Centre. Brown and Farnham labs by James Paris and Mark Takahashi. - [Read Chromatin Immunoprecipitation Protocol]

Category: DNA Microarray Protocols

Chromatin Immunoprecipitation Protocol for Microarray Analysis – Protein A/G Bead Method. Protocol From the UHN Microarray Centre - [Read Chromatin Immunoprecipitation Protocol for Microarray Analysis]

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

Chromatin Immunoprecipitation Protocol for Microarray Analysis – Protein A/G Bead Method. Cross-Linking, Sonication/Chromatin Shearing Efficiency. Reaction UHN Microarray Centre - [Read Chromatin Immunoprecipitation Protocol PDF]

Category: Reporter Gene Assays: Luciferase Assay Protocols

It is possible that some cell lines have lost the ability to perform RNAi or that cells derived from certain tissues do not support RNAi. This reporter assay, for RNAi in mammalian cells, can be used to establish whether the cells under study are susceptible to RNAi. - [Read Cotransfection of Luciferase Reporter Plasmids with siRNA Duplexes Protocol]

Category: Neuroscience Protocols: Neuronal Stains Protocols

An appropriate term for glial fibers is 'nerve glue', because they provide the internal support of the central nervous system.  There are four types of glial cells: astrocytes, oligosendroglia,microglia, and ependymal cells. The glia fibers are stained with crystal violet which are resistant to the aniline-chloroform differentiating solution. - [Read Holzer's Stain Protocol]

Category: Immunology: Mononuclear Cell Isolation Protocols

Presents two methods for preparing dendritic cells (DCs), a highly specialized type of antigen-presenting cell (APC). The first method involves the isolation of DCs from mouse spleen, resulting in a cell population that is highly enriched in accessory cell and APC function. A support protocol for collagenase digestion of splenocyte suspensions is described to increase the yield of dendritic cells. The second method involves generating large numbers of DCs from mouse bone marrow progenitor cells. - [Read Isolation of Dendritic Cells Protocol]

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

Describes flow cytometric protocols using the dyes Indo-1 AM, Fluo-3, and Fura Red AM to measure intracellular calcium concentration. Support protocols detail the use of calcium buffers to calibrate a flow cytometric calcium assay, and methods to facilitate dye loading; an alternate protocol describes the use of a spectrofluorimeter to measure intracellular calcium for those investigators without access to a flow cytometer. - [Read Measurement of Intracellular Ions by Flow Cytometry Protocol]

Category: Chromatography Protocols: Dye Ligand Affinity Chromatography Protocols

Triazine dyes, such as Cibacron Blue 3GA, can be linked to a hexyl spacer arm and then immobilized on a polyhydroxyl support matrix that has been activated with either 1,1-carbonyldiimidazole or epichlorohydrin.  An alternative procedure for immobilizing dyes using the direct coupling method is provided in Immobilization of Dyes on Polyhydroxyl Matrices Using the Direct Coupling Method. - [Read Preparation of Affinity-Ligand Resins by Immobilization of Dyes on Polyhydroxyl Matrices]

Category: Chromatography Protocols: Dye Ligand Affinity Chromatography Protocols

Protocol describes a general approach for the direct attachment of a triazine dye onto a carbohydrate support.  The direct coupling method is achieved under alkaline conditions (pH 10-11) with nucleophilic displacement of the dye’s chlorine atom by the polymer hydroxyls. - [Read Preparation of Affinity-Ligand Resins by Immobilization of Dyes on Polyhydroxyl Matrices Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

A procedure for direct and indirect staining of single-cell suspensions of lymphoid tissue or peripheral blood lymphocytes to detect cell surface membrane antigens is presented. In addition, support protocols present methods for fluorescence labeling of purified antibodies. A protocol for flow cytometric analysis of intracellular antigens in single-cell suspensions is also included. - [Read Preparation of Cells and Reagents for Flow Cytometry Protocols]

Category: Peptide Protocols

Protocol for Manual Peptide Synthesis. Resin Swelling And Coupling Of Activated Amino Acid Esters. Deblocking and cleaving peptide from solid support. Synthesis of Peptides. The University of Nebraska-Lincoln Protein Core Facility - [Read Protocol for Manual Peptide Synthesis]

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

This protocol describes a method for testing of new serum lots prior to use in cell culture. Serum lots vary considerably in their ability to support cell growth, and some lots even contain toxic or growth-inhibitory compounds. It is advantageous to test serum lots and purchase in large volume, both for cost benefit. - [Read Serum Testing for Mammalian Cell Culture Protocol]