western Protocols

Category: Plant Biology Protocols: Arabidopsis Protein Peptide Protocols

Coimmunoprecipitation is one of the most useful techniques for revealing protein-protein interactions. Good negative controls to verify the specificity of the coimmunoprecipitation procedure are (1) performing the same immunoprecipitation experiment using beads coupled to the preimmune serum and (2) probing the Western blot with antibodies against protein known not to interact with the coimmunoprecipitated proteins under physiological conditions. - [Read Protein Coimmunoprecipitation in Arabidopsis Protocol]

Category: Protein Protocols: Protein Trafficking

Methods on protein trafficking. Information on N5 Secretion and protein trafficking. Methods include: Alcian Blue Test; Lucifer Yellow uptake assay; Preparation of total protein extracts for western immunoblots; Screen for Drug Sensitivity. - [Read Protein Trafficking Methods]

Category: Signalling Signal Transduction Protocols

Detection of phosphorylated tyrosine residues can be performed using anti-P-TYR Ab and Western Analysis.Includes 2nd method,which uses phosphotyrosine in conjunction with anti-P-TYR Ab to "unlabel" potential proteins.By comparing Westerns developed with the 1st method(reveals phosphorylated protein) and the 2nd method(reveals non-specific labeling), a more accurate picture of those proteins phosphorylated on tyrosine can be seen. Includes: Protein Preparation, Electrophoresis and Transfer. - [Read Protocol for Antiphosphotyrosine Western Blot Analysis]

Category: Protein Protocols: Western Blot Protocol

Adapted from existing protocols by Vinh PhamLast modified: June 5, 2003. - [Read SDS-PAGE & Western Blotting Protocols]

Category: Protein Protocols: Western Blot Protocol

Simple Western Blotting Protocol ProSci. - [Read Simple Western Blot Protocol]

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

Too many bands on a Western blot FAQs SYSY Synaptic Sys. - [Read Too many bands on a Western blot FAQs SYSY Synaptic Sys.]

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

Troubleshooting Guide for Chemiluminescent Western Blotting Pierce. High Background, Reverse image on film, Blot glows in the darkroom, Signal duration is less than 8 hours, Weak or no signal, Spots within the protein bands, non-specific bands. - [Read Troubleshooting Guide for Chemiluminescent Western Blotting Pierce]

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

Problem: Poor Transfer, Problem: Poor Transfer. R&D Systems. - [Read Troubleshooting Guide: Western Blot]

Category: Protein Protocols: Immunoprecipitation Protocols: Immunoprecipitation Troubleshooting

Troubleshooting the immunoprecipitation procedure Roche. Tagged proteins not visible or only weakly visible on Western blot. High background or many nonspecific bands on gel or blot membrane. - [Read Troubleshooting the immunoprecipitation procedure IP and IB - Roche]

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

Troubleshooting the Western blot procedure Roche. PDF - [Read Troubleshooting the Western blot procedure Roche PDF]

Category: Western Blot Protocols: Antigen Detection Methods Protocols

Use of the chemiluminescence-producing alkaline phosphatase substrate 3-(4-methoxyspiro[1,2-dioxetane-3,2'-tricyclo-[3.3.1.1(3,7)]decan]-4-yl)phenyl phosphate (AMPPD, also known as adamantyl-1,2-dioxetane phosphate), or its dioxetane relatives provides a substantial increase in sensitivity over colorimetric substrates and radiochemical methods currently used for the detection of antigen-antibody complexes immobilized on nylon or PVDF membranes. - [Read Western Analysis Using the Chemiluminescent Alkaline Phosphatase Substrate CSPD Protocol]

Category: Protein Protocols: Western Blot Protocol

Simple Procedure for Western Blotting. Corbett Lab. - [Read Western Blot]

Category: Western Blot Protocols

Procedure describes how it denatures most of the modification and degradation proteins immediately giving the most accurate read out of the true levels of protein at the time of harvest.  However, in cases where detection is a problem, a limited purification (e.g. isolation of nuclear extract for the detection of transcription factors) might be required to allow analysis. - [Read Western Blot Analysis of Endogenous Gene Expression Protocol]

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

This protocol describes nuclear and cytoplasmic fractionation of tissue culture cells and a method for Western blot detection of proteins using the Odyssey Infrared Imaging System. This protocol was used to detect expression of the "small" Tap protein in 293T, HeLa and COS cells. The Odyssey system has several advantages over the more widely used chemiluminescent detection methods. - [Read Western Blot Analysis of Sub-Cellular Fractionated Samples Using the Odyssey Infrared Imaging System]

Category: Immunology: Immunoassays

3-step process of Western immunoblotting for detection of particular sites of phosphorylation on specific proteins recognized by modification-sensitive antibodies. - [Read Western Blot Analysis—Phosphoprotein-Specific Antibody Mixture Protocol]

Category: Protein Protocols: Western Blot Protocol

Protocol includes cell lysate preparation, SDS-PAGE gels, transfer, and blotting. (Bjorkman Group). - [Read Western Blot Detailed (PDF)]

Category: Protein Protocols: Western Blot Protocol

Nice Tips on reducing sample viscosity, buffer preparation, and western blot protocol. Phil Dash. - [Read Western Blot Protocol]

Category: Western Blot Protocols: General Western Blot Methods Protocols

This protocol was developed for the BIORAD protein gel and transfer apparatus.  The buffers can be used with any electrophoresis/transfer system. - [Read Western Blot Protocol]

Category: Protein Protocols: Western Blot Protocol

Theory and Protocol for Western Blotting / immunoblotting. R&D Systems. - [Read Western Blot Protocol and Theory]

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

Streaking of Western Blots, Weak Bands or Weak Staining of Western Blots, Poor Transfer of Membranes, Dirty Blot, Multiple Bands on Western Blot Membrane and Film. Western Blot Info. - [Read Western Blot Troubleshooting]

Category: Buffers Media, and Solutions: Buffer Recipes for Protein Work

Western Blotting and Recipies. Lysis Buffer. - [Read Western Blotting and Recipies]

Category: Protein Protocols: Western Blot Protocol

Western Blotting Using Chemiluminescence. P.J. Hansen. - [Read Western Blotting Using Chemiluminescence]

Category: Protein Protocols: Western Blot Protocol

Detailed protocol for Western Blot using Biotinylated Antibodies. (BD PharMingen). - [Read Western Blotting with Biotinylated Antibodies]

Category: Protein Protocols: Western Blot Protocol

Detailed General Protocol for Western Blotting with Monoclonal Antibodies. PharMingen - [Read Western Blotting with Monoclonal Antibodies]

Category: Yeast Protocols: Yeast Protein Protocols

Protocol is a simple, reliable method for the preparation of yeast protein extracts for analysis by polyacrylamide gel electrophoresis (PAGE) and Western blotting. - [Read Yeast Protein Extracts Protocol]