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Protein Protocols MolecularStation - http://www.molecularstation.com/protein/protein-protocols/

Protein Protocols MolecularStation. Western Blotting, Immunoprecipitation, stripping and reprobing - [Read Protein Protocols MolecularStation]

Protein and Antibody Microarray Chips - An Introduction - http://www.molecularstation.com/protein-microarrays/

Category: Protein Microarray Protocols

Protein Microarray Chips - An Introduction. Introduction, Types of Protein Chips, Attachment, Protein and Antibody Chip Production, Applications of Protein Chips, Detection methods, and Future Directions. Molecularstation. - [Read Protein and Antibody Microarray Chips - An Introduction]

Western Blot Home - http://www.molecularstation.com/protein/western-blot/

Category: Protein Protocols: Western Blot Protocol

Western Blot Home. Information on western blotting, western blot procedure and methods, western blot books, stripping protocols. - [Read Western Blot Home]

Troubleshooting Western Blot - http://www.molecularstation.com/protein/troubleshooting-western-blots/

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

Covers Many Western Blot Problems and Includes Many Solutions. Fuzzy Bands, Low or Weak Signal, High Background, Spots on Film, Too Many Bands. A MolecularStation Guide. - [Read Troubleshooting Western Blot]

Search Results Protocol Links Sort by: Hits | Alphabetical

Gel Retardation Assays for DNA-binding Proteins Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3948

Category: Nucleic Acid Electrophoresis Protocols: DNA Electrophoresis Protocols: DNA Electrophoresis in Polyacrylamide Gels Protocols

Protocol exploits differences in electrophoretic mobility through a nondenaturing polyacrylamide gel between a rapidly migrating target DNA and a more slowly migrating DNA-protein complex. - [Read Gel Retardation Assays for DNA-binding Proteins Protocol]

Gel Super-Shift Assay EMSA Protocol - http://www.biochem.northwestern.edu/ibis/morimoto/research/Protocols/III.%20Proteins/C.%20DNA-Protein%20Interactions/3b.%20Super-shift.pdf

Category: DNA Protein Interactions Protocols: Electrophoretic Mobility Shift Assays EMSA Protocols

Protocol for gel super shift assay EMSA. - [Read Gel Super-Shift Assay EMSA Protocol]

Gene Expression and Protein Localization in Arabidopsis Protocols - http://www.arabidopsis.org/cshl-course/7-gene_expression.html

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocols for gene expression and protein localization in Arabidopsis. Includes: Detection of the native protein; Detection of a recombinant version; Immunofluorescence detection in Arabidopsis protoplasts; Isolation of Arabidopsis seedling protoplasts; Subcellular localization of GUS-fusion proteins in Arabidopsis seedlings; Localization of Arabidopsis proteins with GUS in situ enzyme assay. - [Read Gene Expression and Protein Localization in Arabidopsis Protocols]

Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4562

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Protein Protein Interactions Protocols

This procedure capitalizes on the asexual/sexual reproductive cycle of yeast cells. Laboratory yeast strains can be maintained indefinitely by asexual reproduction as one of two haploid mating types, MATa or MAT{alpha}. - [Read Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array]

Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array: - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4563

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Protein Protein Interactions Protocols

This protocol describes how to handle arrayed yeast using robotics. - [Read Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array:]

Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array: Amplification of ORFs - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4561

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Protein Protein Interactions Protocols

This protocol describes the first step in constructing an array: amplification of the predicted ORFs that are to be included in the array. Gene-specific primers containing vector-specific flanking sequences that facilitate recombinational cloning are used to amplify each ORF. A secondary amplification can be used to extend the length of the homologous vector sequence flanking the ORF. - [Read Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array: Amplification of ORFs]

Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array: S - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4564

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Protein Protein Interactions Protocols

This protocol describes three different storage methods for the array. Frozen stocks are the most permanent method and are an absolute requirement for maintaining a record of the array. - [Read Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array: S]

GST Fusion Protein Prep - http://cmgm.stanford.edu/~vesicle/protocols/gst_prot.html

Category: Molecular Model Organisms: Yeast Two-Hybrid Screen Protocol

Detailed protocols and methods for GST protein purification. Vesicle Trafficking, Stanford. - [Read GST Fusion Protein Prep]

GST Fusion Protein Purification from Yeast - http://www.med.unc.edu/~hdohlman/gst.html

Category: Molecular Model Organisms: Yeast Two-Hybrid Screen Protocol

GST Fusion Protein Purification from Yeast. Lysis Buffer recipe for yeast, glutathione bead method for GST purification. Dohlman Lab - [Read GST Fusion Protein Purification from Yeast]

GST Fusion Protein Purification from Yeast Protocol - http://www.med.unc.edu/~hdohlman/gst.html

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

Protocol for GST fusion protein purification from Yeast. - [Read GST Fusion Protein Purification from Yeast Protocol]

GTPÎ³S-Induced Inhibition of cAMP Binding to the cAMP Receptor (cAR1) in Dictyostelium Discoideum - http://www.bio.com/protocolstools/protocol.jhtml?id=p1175

Category: Signalling Signal Transduction Protocols

This protocol assays inhibition of in vivo binding of [3H]-cAMP to cAR1 by GTPÎ³S. Dictyostelium discoideum respond to extracellular cAMP through the cAMP chemoattractant receptor (cAR1). Binding of cAMP to the G protein-coupled cAR1 is inhibited by the GTP analog GTPÎ³S. Protocol includes information on: Solutions used, BioReagents and Chemicals and Protocol Hints. - [Read GTPÎ³S-Induced Inhibition of cAMP Binding to the cAMP Receptor (cAR1) in Dictyostelium Discoideum]

GUS Reporter Gene Assay Protocol - http://www.oardc.ohio-state.edu/stockingerlab/Protocols/GUS%20Assays.pdf

Category: Reporter Gene Assays

Protocol for GUS reporter gene assay. Includes: Protein isolation; Alternative method for small (<1g) quantities of tissue; GUS assays; Bradford protein concentration determination assays - [Read GUS Reporter Gene Assay Protocol]

High Yield Protein Production from Pichia pastoris Yeast PDF - http://www.nbsc.com/files/papers/abl_pichia.pdf

Category: Protein Protocols: Protein Expression Protocols

High Yield Protein Production from Pichia pastoris Yeast PDF. Julia Cino, PhD - [Read High Yield Protein Production from Pichia pastoris Yeast PDF]

His Tag Nickel Affinity Chromatography Protocol PDF - http://www.mnstate.edu/provost/His_TagNiChromatogProtocol.pdf

Category: Chromatography Protocols

His Tag Nickel Affinity Chromatography Protocol PDF. The Wallert and Provost Lab. Theory and Introduction: Ni-Affinity Chromatography uses the ability of His to bind nickel. Six histadine amino acids at the end of a protein (either N or C terminus) is known as a 6X His tag. Nickel is bound to an agarose bead by chelation using nitroloacetic acid (NTA) beads. Several companies produce these beads as His Tagged proteins are some of the most used affinity tags in todayâ€™s market. - [Read His Tag Nickel Affinity Chromatography Protocol PDF]

How to Isolate Nondenatured Protein Extracts from Neurospora for Western Analysis - http://www.fgsc.net/neurosporaprotocols/How%20to%20isolate%20nondenatured%20protein%20extracts%20from%20Neurospora%20for%20western%20analysis.pdf

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]

Identification of Associated Proteins by Coimmunoprecipitation Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3898

Category: Immunology: Immunoprecipitation Protocols: Immunoprecipitation of Radiolabeled Cells Protocols

Coimmunoprecipitation is most commonly used to test whether two proteins of interest are associated in vivo, but it can also be used to identify novel interacting partners of a target protein. In both cases, the cells, which may have been labeled with [35S]methionine, are harvested and lysed under conditions that preserve protein-protein interactions. The target protein is specifically immunoprecipitated from the cell extracts, and the immunoprecipitates are fractionated by SDS-PAGE. - [Read Identification of Associated Proteins by Coimmunoprecipitation Protocol]

Immunoaffinity Purification: Coupling Antibodies to Protein A or G Bead Columns Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4303

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

Protocol describes how antibodies are bound to protein A or protein G beads and are then crosslinked to the matrix via a bifunctional coupling reagent called dimethyl pimelimidate (DMP). - [Read Immunoaffinity Purification: Coupling Antibodies to Protein A or G Bead Columns Protocol]

Immunoblot Protocol - http://cgap-mf.nih.gov/Protocols/DNARNAProteomicAnalysis/Proteomics/Immunoblot.html

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Protocol for Immunoblot. Includes: Staining and Laser Capture Microdissection; Protein Separation by Polyacrylamide Gel Electrophoresis; Electrophoretic Transfer To a Membrane (Nylon, PVDF or Nitrocellulose); Primary and Secondary Antibody Incubations; Visualization. - [Read Immunoblot Protocol]

Immunoblotting: Preparing Immunoprecipitated Proteins Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4300

Category: Western Blot Protocols

Protein immunoprecipitation can be a useful preparative step for immunoblotting. For very rare proteins, the protein of interest can be purified and concentrated by standard immunoprecipitation techniques before immunoblotting. In addition, protein-protein interactions can be tested with an immunoprecipitating antibody that is specific for one protein of a complex and an immunoblotting antibody that is specific for a second member of the complex. - [Read Immunoblotting: Preparing Immunoprecipitated Proteins Protocol]

Immunoblotting: Preparing Protein Solutions Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4298

Category: Western Blot Protocols: General Western Blot Methods Protocols

Protocol describes how proteins are mixed with a sample buffer and prepared for electrophoresis on standard Tris/glycine SDS-polyacrylamide gels. - [Read Immunoblotting: Preparing Protein Solutions Protocol]

Immunofluorescence Detection of RNAi-Induced Protein Knockdown in Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4344

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]

Immunofluorescence Detection of RNAi-Induced Protein Knockdown in Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4344

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]

Immunoprecipitation of mRNA-Protein Complexes Protocol - http://www.nature.com/nprot/journal/v1/n2/pdf/nprot.2006.82.pdf

Category: Immunology: Immunoprecipitation Protocols

Protocol for immunoprecipitation of mRNA-protein complexes. In this protocol, an antibody targeting an RBP of interest is used to immunoprecipitate the RBP and any interacting molecules from a cell lysate. Reverse transcription followed by PCR is then used to identify individual mRNAs isolated with the RBP. This method focuses on examining an association between a specific RBP-mRNA complex, and it is best suited for a small scale screening of known or putative binding partners. - [Read Immunoprecipitation of mRNA-Protein Complexes Protocol]

Immunoprecipitation of Proteins Method - IP protocol - http://www.upstate.com/misc/protocols.q.prot.e.immuno/Immunoprecipitation+of+Proteins

Category: Protein Protocols: Immunoprecipitation Protocols

Ip Protocol using protein A or G beads. Upstate. - [Read Immunoprecipitation of Proteins Method - IP protocol]

Immunoprecipitation: Denaturing Lysis Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4534

Category: Immunology: Immunoprecipitation Protocols

For many sources of antigens, one useful method of lysis is to treat cells with harsh, denaturing solutions to release most of the protein antigens, as described here. The lysates are then diluted to reduce the denaturing conditions to levels that are suitable for the formation of antibody-antigen complexes. The resulting solution is precleared prior to immunoprecipitation. - [Read Immunoprecipitation: Denaturing Lysis Protocol]

Articles (1-10 of 10)

Populational Analysis of Genomic DNA Protocol

A protocol on the populational Analysis of genomic DNA.

Lambda Phage Protocol for Large Preparation DNA Recovery

Lambda Phage Protocol for Large Preparations DNA Recovery.

Acid Guanidinium Thiocyanate RNA Isolation Phenol Chloroform Extraction

A single step RNA isolation protocol using Phenol Chloroform Extraction and Acid Guanidinium Thiocyanate. This RNA isolation method uses the fact that guanidinium thiocyanate can simultaneously lyse the cells and inactive cellular RNAses during the initial RNA isolation step allow a single step in the method.

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.

In Vitro Translated Xenopus Mos Kinase Assay Protocol

In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.

Antibodies Phage Expression Protocol Using 96-well Microtiter Plates

A Phage display expression protocol for antibody expression in 96-well microtiter plates.

Immobilized Antigen Phage Antibody Selection Protocol

A protocol for the selection of Phage Antibodies using Immobilized Antigen. This method describes the selection of antibodies from bacteriophage antibody libraries that recognize a specific antigen. The phage display library of antibody-displaying phage particles is exposed to antigen attached to a solid substrate (Nunc Immuno™ tubes). The phage particles with affinity for antigen bind to the immobilized antigen and are selected from the library of phage expressing antibodies.

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