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Antibody Addition to Drosophila Specimens and Detection Using Enzyme-Linked Reagents Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4526

Category: Drosophila Protocols

Enzyme-linked reagents give excellent sensitivity and use a simple light microscope for detection. A range of enzymes is available, but for staining in situ, horseradish peroxidase will suit most needs. Diaminobenzidine (DAB) is one of the most sensitive substrates for horseradish peroxidase. It yields an intense brown product that is insoluble in both water and alcohol. It can be made more sensitive by adding metal salts such as cobalt or nickel to the substrate solution. - [Read Antibody Addition to Drosophila Specimens and Detection Using Enzyme-Linked Reagents 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]

Construction of Micro-Tip for Use in IMAC Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000163.pdf?pid=PP00000163

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Protocol is the second step in a three-step process for the preparation and enrichment of phosphopeptides using immobilized metal affinity chromatography (IMAC) for the identification of the phosphopeptides by LCMS/ MS. This procedure describes the construction of microchromatographic columns, or micro-tips. - [Read Construction of Micro-Tip for Use in IMAC Protocol]

Lysis and Protein Extraction from WEHI-231 Cells with TriPure Isolation Reagent Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000162.pdf?pid=PP00000162

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Protocol is the first step in a three-step process for the preparation and enrichment of phosphopeptides using immobilized metal affinity chromatography (IMAC) for the identification of the phosphopeptides by LC-MS/MS. This procedure is used to prepare protein extracts from WEHI-231 cells. This preparation method provides total cellular protein samples that are free of contaminating nucleic acids. - [Read Lysis and Protein Extraction from WEHI-231 Cells with TriPure Isolation Reagent Protocol]

Optimization of Imidazole Concentrations for Immobilized Metal-Ion Affinity Chromatography Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/9/pdb.prot4261

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Protocol for the optimization of imidazole concentrations for immobilized metal-ion affinity chromatography. Most samples from which histidine-tagged proteins are to be purified also contain endogenous protein contaminants that bind to the immobilized metal-ion affinity chromatography (IMAC) adsorbent. Usually, these proteins bind more weakly than the histidine-tagged protein. - [Read Optimization of Imidazole Concentrations for Immobilized Metal-Ion Affinity Chromatography Protocol]

Optimized Protocol for Mounting Tissue Sections PDF - http://arctur.com/research_portal/images/Application%20Notes/pen_membrane_protocol_9.pdf

Category: Histology Protocols: Tissue Sample Sectioning

Generally Required Materials and Equipment for Tissue Sectioning. Sectioning and Slide Preparation Procedure onto Metal-Framed PEN Membrane Slides. Arcturus. - [Read Optimized Protocol for Mounting Tissue Sections PDF]

Packing an IMAC Column Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4215

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Protocol for packing an IMAC column. Protocol describes a procedure for packing a 30-ml immobilized metal-ion affinity chromatography (IMAC) column with the aid of a pump. - [Read Packing an IMAC Column Protocol]

Preparation and Enrichment of Phosphopeptides from Phosphotyrosine Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000193.pdf?pid=PP00000193

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Protocol is specifically for the further enrichment of phosphopeptides from a phosphotyrosine pull-down. This is the final step for the preparation and enrichment of phosphopeptides using immobilized metal affinity chromatography (IMAC) for the identification of the phosphopeptides by liquid chromatography tandem mass spectrometry (LC-MS/MS). - [Read Preparation and Enrichment of Phosphopeptides from Phosphotyrosine Protocol]

Preparation and Enrichment of Phosphopeptides Using IMAC and LC-MS/MS - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000164.pdf?pid=PP00000164

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Protocol is the third step in a three-step process for the preparation and enrichment of phosphopeptides using immobilized metal affinity chromatography (IMAC) for the identification of the phosphopeptides by LCMS/ MS. - [Read Preparation and Enrichment of Phosphopeptides Using IMAC and LC-MS/MS]

Preparation of a Prepacked IMAC Column Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4214

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Protocol for the preparation of a prepacked IMAC column. Purification of histidine-tagged proteins on a milligram scale can be performed conveniently on a small (1to 5-ml) immobilized metal-ion chromatography (IMAC) column using a syringe to load the sample, wash the column, and elute the protein. - [Read Preparation of a Prepacked IMAC Column Protocol]

Protein Purification Using Immobilized Metal-Ion Affinity Chromatography under Optimized Imidazole - http://www.cshprotocols.org/cgi/content/extract/2006/9/pdb.prot4262

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Perform this protocol after determining the imidazole concentration that yields the best purification of the recombinant protein of interest. - [Read Protein Purification Using Immobilized Metal-Ion Affinity Chromatography under Optimized Imidazole]

Purification of Histidine-Tagged Proteins under Denaturing Conditions Using IMAC Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4221

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Immobilized metal-ion affinity chromatography (IMAC) is suitable for the purification of proteins under denaturing conditions. Either guanidine-HCl or urea can be used, although guanidine-HCl is a stronger denaturant than urea. Proteins that have been adsorbed to the column in the presence of guanidine-binding buffer may be washed with urea-binding buffer and eluted with urea elution buffer. - [Read Purification of Histidine-Tagged Proteins under Denaturing Conditions Using IMAC Protocol]

Purification of Histidine-Tagged Proteins Using IMAC Without Parameter Optimization Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4217

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Histidine-tagged proteins can be purified on prepacked 1-ml immobilized metal-ion affinity chromatography (IMAC) columns without optimization of the separation conditions. The method allows fast capture of the target protein, although with a lower purity than can be obtained under optimized conditions. - [Read Purification of Histidine-Tagged Proteins Using IMAC Without Parameter Optimization Protocol]

Staining of Nerve Fibers Protocol - http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/BODIAN.PDF

Category: Neuroscience Protocols: Neuronal Stains Protocols

Protargol-S (silver proteinate) is used with the addition of copper metal. The copper replaces the silver in the connective tissue, allowing a greater differentiation between the nerve fibers and the connective tissue. The silver is reduced with hydroquinone to the visible metallic form. The sections are toned with gold chloride, the gold chloride is reduced with oxalic acid, increasing the deposit of metallic gold on the sections. - [Read Staining of Nerve Fibers Protocol]

Unmasking Hidden Epitopes Using the Pressure Cooker Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4530

Category: Immunohistochemistry Protocols: Antigen Retrieval Protocols

The principle behind the pressure cooker method described here is to use extended periods of heat to break some of the subcellular structures that block antibody access. This approach is appropriate for handling specimens on glass slides. The major advantages of the pressure cooker method are the ability to handle a large number of slides simultaneously, the convenience of using metal racks, and the avoidance of any hot spots that are found in the microwave. - [Read Unmasking Hidden Epitopes Using the Pressure Cooker Protocol]

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Paraffin Embedding Protocol

Paraffin Embedding Protocol for molecular profiling. This Paraffin Embedding Protocol describes the processing of the tissues into sections following ethanol fixation. Molecular profiling (MP) is a technique that is used to visualize the global patterns of RNA expression or protein expression in various cell types and disease processes.