western Protocols

Category: Protein Protocols: Western Blot Protocol

Cheap chemiluminescent detection for western blotting. Yakunin AF 1998. - [Read Almost Homemade Chemiluminescent Detection]

Category: Protein Protocols: Protein Electrophoresis

An excellent guide on the analysis of proteins on SDS-PAGE gels, through staining with coomassie blue dye and western blot analysis. Analysis of Protein Gels (SDS-PAGE). David R. Caprette, Rice University. - [Read Analysis of Protein Gels (SDS-PAGE)]

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Protein Purification and Isolation Protocols

Preparation of protein extracts for western blot. - [Read Boiling Protein Prep Protocol]

Category: Protein Protocols: Western Blot Protocol

How to prepare cell lysates for western blotting. - [Read Cell Lysates For Western Blotting]

Category: Signalling Signal Transduction Protocols: Chemokine Protocols

Western Blotting chemokine receptors, Immunohistochemistry on Paraffin sections for chemokines. - [Read Chemokine and Interleukin Antibody Protocols]

Category: RNAi Technology Protocols: RNAi Mammalian Cells Protocols

Protocol describes the use of western blotting to analyze the RNAi-induced depletion of target protein from mammalian cells.  Immunofluorescence can be used as an alternative. - [Read Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting Protocol]

Category: Western Blot Protocols

Protocol describes the use of western blotting to analyze the RNAi-induced depletion of target protein from mammalian cells.  Immunofluorescence can be used as an alternative. - [Read Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting Protocol]

Category: Yeast Protocols: Yeast Protein Protocols

This assay is performed to detect ubiquitylated proteins in yeast. Yeast that have been transformed with a vector expressing polyhistidine-tagged ubiquitin (Ub) under the control of a copper-inducible promoter are grown, induced with copper, and harvested. Total ubiquitylated proteins are then recovered by nickel-affinity chromatography, and specific proteins are detected by Western blotting. - [Read Detection of Ubiquitylated Proteins in Yeast Protocol]

Category: Protein Protocols: Western Blot Protocol

ENZYME-ASSISTED IMMUNOELECTROBLOTTING (IEB OR WESTERN BLOTTING. E.P. Rybicki and Maud Purves. Department of Microbiology. Copper Staining of Gels, Indirect enzyme immunoassay, Staining Proteins on Membranes, Blotting Buffer, Staining of proteins in gels - [Read ENZYME-ASSISTED IMMUNOELECTROBLOTTING (IEB OR WESTERN BLOTTING)]

Category: Immunology: Immunoprecipitation Protocols

Protocols for detection and purification of proteins tagged with a particular epitope, the FLAG tag, although the same general approach can be applied to other epitope tags. The protocols employ the anti-FLAG M2 antibody to detect and purify FLAG-tagged proteins. The methods presented are immunoprecipitation of FLAG fusion proteins from cells using an anti-FLAG M2 affinity gel, detection of FLAG fusion proteins by western blotting, and purification of FLAG fusion proteins by anti-FLAG. - [Read Epitope Tagging of Recombinant Proteins Protocol]

Category: Cell Biology and Cell Culture: Cell Culture Techniques: Cell Line Preservation Freezing

Freezing and Thawing Eukaryotic Cells in liquid nitrogen, Freezing Cell Lysates for Western Blots, Thawing Cells from Liquid Nitrogen. Dr. Bart Frank Oklahoma. - [Read Freezing and Thawing Eukaryotic Cells]

Category: Signalling Signal Transduction Protocols

General protocol for Ras, Rac, Cdc42, and Rho activation assay. Includes: Affinity Precipitation/Immunoblot Protocol, Cell Culture and Extract Preparation (Adherent and Non Adherent cells), GTPγS/GDP Loading for Positive and Negative Controls, Ras, Rac ,Cdc42, and Rho Pull-Down Assay and Western Blot and Detection. - [Read General Method for Ras, Rac, Cdc42, and Rho Activation Assay]

Category: Western Blot Protocols

Protocol for high throughput western blotting for antibody quality control. Includes: SDS-PAGE; TRANSFER; IMMUNOSTAINING; DETECTION. - [Read High Throughput Western Blotting Protocol for Antibody Quality Control]

Category: Protein Protocols: Western Blot Protocol

All the products and steps to complete a full western blotting analysis. (Jonathan Flint lab). - [Read How to do a Western Blot]

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to isolate nondenatured protein extracts from Neurospora for western analysis. - [Read How to Isolate Nondenatured Protein Extracts from Neurospora for Western Analysis]

Category: Protein Protocols: Western Blot Protocol

Nice Western Blotting Protocol using some eBioscience Products. - [Read Immunoblotting Using eBioscience Products]

Category: Immunology: Immunofluorescence Protocols

This protocol describes the use of a specific antibody that recognizes the targeted gene product to detect RNAi-induced gene knockdown in mammalian cells. Western blot technology can be used as an alternative (see Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting). - [Read Immunofluorescence Detection of RNAi-Induced Protein Knockdown in Mammalian Cells Protocol]

Category: RNAi Technology Protocols: RNAi Mammalian Cells Protocols

Protocol describes the use of a specific antibody that recognizes the targeted gene product to detect RNAi-induced gene knockdown in mammalian cells.  Western blot technology can be used as an alternative. - [Read Immunofluorescence Detection of RNAi-Induced Protein Knockdown in Mammalian Cells Protocol]

Category: Immunology: Immunoprecipitation Protocols

Protocol for immunoprecipitation. Includes: Preparing Cell Lysate; Immunoprecipitating; Western Blotting. - [Read Immunoprecipitation Protocol II]

Category: Western Blot Protocols: Antigen Detection Methods Protocols

Protocol describes, samples containing the target protein are deposited onto a polyvinyldifluoride (PVDF) membrane using a vacuum manifold.  The immobilized protein is exposed to an antibody specific for the target protein, followed by an antibody that reacts with species-specific determinants carried by the primary antibody and is conjugated to horseradish peroxidase (HRP). - [Read Measuring Protein Concentration by Western Analysis Using Enhanced Chemiluminescence Detection]

Category: Protein Protocols: Western Blot Protocol: Troubleshooting Western Blots

Optimizing Western Blots: Pre-Transfer Invitrogen - [Read Optimizing Western Blots: Pre-Transfer Invitrogen]

Category: Antibody Protocols: Antibody Production Protocols

Protocol for polyclonal antibody production. Very useful for rapid and simple generation of antibodies for western blots, ELISA assays, and immunoprecipitation. Includes: Rabbit Immunization; Initial Preparation; Pre-bleed; Antigen Injection; Monitoring of Titer; Purification of Antibodies. - [Read Polyclonal Antibody Production Protocol]

Category: Protein Protocols: Western Blot Protocol

Western blotting using enriched membrane preparation and postsynaptic densities are desired. (BD Biosciences) - [Read Preparation of Brain Membrane Fractions for Western Blot Analysis]

Category: Protein Protocols: Nuclear Extraction Protocols

Nuclear Extract protocol is optimized for about 10^7 cells, can use for gel shift assays and western blotting. Mirmira Lab, Univ. Virginia - [Read Preparation of Nuclear Extracts for Gel Shifts and Westerns Blots]

Category: C. elegans Protocols

Protocol was designed to rapidly generate small scale cytosolic extracts of C. elegans for Western or IP (has not been tested for RNA work). The protocol works well for between 50 to 5000 worms and has not been extensively tested on larger a scale, though it should work. Includes: Collection; Sonication; Clearing lysate; Immunoprecipitation. - [Read Preparation of Worm Extracts by Sonication Protocol]