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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

Lysing Tissue-Culture Cells for Immunoprecipitation Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4531

Category: Cell Culture Protocols: Cell Lysis Protocols

For cells grown in tissue culture, the most useful method of lysis is treating with detergents, as described in this protocol. Non-ionic detergents, such as NP-40, solubilize the plasma and intracellular membranes, break many weak intermolecular bonds, and solubilize most of the commonly studied protein antigens. RIPA lysis buffer may be used as a more rigorous extraction buffer to release all but the insoluble proteins of the cell and to break most weak noncovalent interactions. - [Read Lysing Tissue-Culture Cells for Immunoprecipitation 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]

Maintaining Live Cells on the Microscope Stage - http://www.microscopyu.com/articles/livecellimaging/livecellmaintenance.html

Category: Microscopy Protocols: Live-Cell Imaging Protocols

Live-cell imaging techniques provide a critical insight into the fundamental nature of cellular and tissue function, especially due to the rapid advances that are currently being witnessed in fluorescent protein and synthetic fluorophore technology. Because of these advances, live-cell imaging has become a requisite analytical tool in most cell biology laboratories. - [Read Maintaining Live Cells on the Microscope Stage]

Mapping Protein-binding Sites on DNA by Dnase I Footprinting Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3947

Category: DNA Protein Interactions Protocols: Dnase Footprinting Assay Protocols

Dnase I is used to fragment a radiolabeled target DNA in the presence and absence of a nuclear extract. A "footprint" is generated when a protein binds to the target and protects a specific segment of DNA from the nucleolytic activity of Dnase I. By comparing the electrophoretic mobility of the Dnase I cleavage products to those of a sequence ladder derived from the same DNA fragment, the position(s) of the DNA sequences recognized by DNA-binding proteins can be determined. - [Read Mapping Protein-binding Sites on DNA by Dnase I Footprinting Protocol]

Mass spectrometry analysis and identification of proteins - http://www.healthsystem.virginia.edu/internet/biomolec/mass_spectrometry.cfm

Category: Mass Spectrometry Protocols

Protein sequencing by mass spectrometry. Mass analysis of peptides and proteins. Identification of modifications in proteins. Good information.Univ. Virginia. Dr. Nicholas Sherman - [Read Mass spectrometry analysis and identification of proteins]

Measurement of Green Fluorescent Protein Expression and DNA Content in Unfixed Cells Protocol - http://cyto.mednet.ucla.edu/Protocols/GFPandDNAHoechst.htm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Data Analysis Protocols

Protocol for measurement of green fluorescent protein expression and DNA content in unfixed cells. - [Read Measurement of Green Fluorescent Protein Expression and DNA Content in Unfixed Cells Protocol]

Measuring Protein Concentration by Western Analysis Using Enhanced Chemiluminescence Detection - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4255

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]

Mechanical Tracing of Protein Function in the Drosophila Ear Protocol - http://www.natureprotocols.com/2006/11/03/mechanical_tracing_of_protein.php

Category: Drosophila Protocols

Method relies on the examination of the ear’s mechanics, which is actively modulated by the motility of auditory neurons and reflects the function of mechanosensory proteins these cells comprise [5-7]. Mechanical signatures arising from the motility of the neurons are assayed by measuring the vibrations of the antennal sound receiver in the presence and absence of sound. - [Read Mechanical Tracing of Protein Function in the Drosophila Ear Protocol]

Metabolic Labeling & Immunoprecipitation Protocols - http://www.cbrinstitute.org/labs/springer/protocols/jun_35Slabeling_IP.html

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

Cells incorporate 35S-methionine or cysteine during the protein synthesis. Thus it is essential to use Met,Cys-free medium and dialyzed FCS during the labeling. Short period of incubation with 35S-methionine or cysteine will result in radiolabeling (pulse), and additional incubation with excess concentration of unlabeled Met+Cys (chase) is needed for complex glycoproteins like integrins to get expressed as a maturated form. - [Read Metabolic Labeling & Immunoprecipitation Protocols]

Microdissection Overview - http://cgap-mf.nih.gov/Protocols/Microdissection/Overview.html

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

Human tissues are comprised of multiple interacting cell populations in a complex three dimensional arrangement with each cellular phenotype determined by a unique profile of mRNA and protein expression. Before microdissection techniques were developed, the only analysis tools for phenotypic studies were primarily immunohistochemistry and in-situ hybridization. While useful, these tools are limited to single gene analysis and, in general, do not allow qualitative studies. - [Read Microdissection Overview]

Microinjection of Protein Samples Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4657

Category: Microinjection Protocols: Microinjection of Proteins Protocols

This protocol describes a method for microinjecting proteins into the nucleus or cytoplasm of adherent cells. Microinjection equipment can be obtained from a number of suppliers; this protocol has been used with the Narishige IM-200 air pressure regulator and the Leitz micromanipulator. Using this system, it is possible to microinject a constant volume within a 50% difference among cells. - [Read Microinjection of Protein Samples Protocol]

Millipore's ZipTip Protocol - http://www.millipore.com/publications.nsf/docs/tn072

Category: Mass Spectrometry Protocols

Millipore's ZipTip Protocol. Intended for purifying and concentrating femtomoles to picomoles of protein, peptide or oligonucleotide samples prior to analysis, providing better data quality. Millipore. - [Read Millipore's ZipTip Protocol]

Mitogen Activated Protein Kinase Guide - http://www.promega.com/guides/sigtrans_guide/strg_chap01.pdf

Category: Signalling Signal Transduction Protocols: MAPK Protocols

Mitogen Activated Protein Kinase Guide. MEK, MEKK, MAPK, MAPKK, antibodies and kits Promega. - [Read Mitogen Activated Protein Kinase Guide]

Mixed Oligonucleotide-primed Amplification of cDNA MOPAC Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3484

Category: PCR Protocols: cDNA Synthesis Protocols

In MOPAC, the amino-terminal and carboxy-terminal sequences of a peptide are used to design two redundant families of oligonucleotides encoding the aminoand carboxy-terminal sequences of the peptide. The primers are used either to amplify a segment of cDNA prepared by RT-PCR from a tissue known to express the protein or to amplify a segment of DNA from an established genomic or cDNA library. - [Read Mixed Oligonucleotide-primed Amplification of cDNA MOPAC Protocol]

Monitoring Protein Phosphorylation for the Ligand Screens PPT - http://www.signaling-gateway.org/aboutus/supplement/AM2003/10a_Mumby.ppt

Category: Protein Protocols: Protein Phosphorylation Protocols

A powerpoint slide demonstrating a method for Monitoring Protein Phosphorylation for Ligand Screens. Mumby. signaling-gateway.org - [Read Monitoring Protein Phosphorylation for the Ligand Screens PPT]

Monster Green® Fluorescent Protein Assay - http://www.promega.com/catalog/country_select.asp?/tbs/tb320/tb320.html&ckt=2

Category: Reporter Gene Assays: GFP Assay Protocols

Green fluorescent protein is commonly used to monitor gene expression and protein trafficking within intact cells. The Monster Green® Fluorescent Protein is encoded by an improved synthetic version of the green fluorescent protein gene originally cloned from Montastrea cavernosa (Great Star Coral). - [Read Monster Green® Fluorescent Protein Assay]

Neutral Red Cytotoxicity Assay Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/64.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

The cytotoxic effect of chemicals upon mammalian cells, such as BALB/c 3T3 and HepG2, in culture is measured by highest tolerated dose (HTD), cell viability (Neutral Red) and total cell protein (coomassie blue). - [Read Neutral Red Cytotoxicity Assay Protocol]

No Boil Yeast Extracts Protocol - http://www.ciwemb.edu/labs/koshland/Protocols/Protein/noboil.html

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

Protocol for no boil Yeast extracts. - [Read No Boil Yeast Extracts Protocol]

Nondenaturing Protein Immunoprecipitation from Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4620

Category: Immunology: Immunoprecipitation Protocols: Protein Immunoprecipitation Protocols

Protocol describes a nondenaturing immunoprecipitation (IP) for mammalian cells. Prefer to use denaturing IPs to recover labeled proteins from pulse-chase experiments. However, the nondenaturing protocol is useful when one wishes to separate soluble from insoluble proteins, or when the antibody being used recognizes a native epitope. - [Read Nondenaturing Protein Immunoprecipitation from Mammalian Cells 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 Protocols for Fluorescent in situ Hybridization in Drosophila Tissues - http://www.utoronto.ca/krause/documents/FISHCHAPTER.pdf

Category: Drosophila Protocols: Drosophila Genetics Protocols

Optimized protocols for fluorescent in situ hybridization in Drosophila tissues. Includes: RNA Probe Preparation; Initial Embryo Fixation; Post-Fixation, Hybridization and post-Hybridization Washes; Development of FISH Signal; Mounting and Viewing of Samples; Double FISH; FISH on Dissected Tissues; RNA-Protein Double-labeling. - [Read Optimized Protocols for Fluorescent in situ Hybridization in Drosophila Tissues]

Overview of Affinity Purification in Combination with Mass Spectrometry Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/30/pdb.top3

Category: Chromatography Protocols: Affinity Chromatography Protocols

Protein complexes can be isolated by several different approaches. For example, a protein can be tagged with an epitope such as Flag or TAP and then overexpressed in a target cell, allowing the interacting proteins to be purified. Similarly, epitope tags can be homologously recombined into the endogenous locus ("knocked-in"), allowing protein complexes containing the tagged proteins to be isolated at their natural expression level. - [Read Overview of Affinity Purification in Combination with Mass Spectrometry Protocol]

PERSPECTIVES ON BACULOVIRUS EXPRESSION SYSTEMS - http://omrf.ouhsc.edu/~frank/baculsum.html

Category: Recombinant Protein Protocols: Baculovirus Recombinant Protein

PERSPECTIVES ON BACULOVIRUS EXPRESSION SYSTEMS. Advantages and disadvantages of protein expression in baculovirus . Insect Cell Culture. Frank Lab. - [Read PERSPECTIVES ON BACULOVIRUS EXPRESSION SYSTEMS]

Ph.D. Peptide Display Cloning System - NEB - http://www.neb.com/nebecomm/TechBulletinFiles/techbulletinE8101.pdf

Category: Peptide Protocols: Phage Display Cloning Systems and Protocols

Ph.D. Peptide Display Cloning System - NEB. The Ph.D.â„¢ Peptide Display Cloning System facilitates the display of custom peptide libraries on the surface of bacteriophage M13 as coat protein fusions, creating a physical linkage between each displayed pep - [Read Ph.D. Peptide Display Cloning System - NEB]

Pierce Protein Glycosylation Kits - http://www.piercenet.com/Products/Browse.cfm?fldID=7CE3FCF5-0DA0-4378-A513-2E35E5E3B49B

Category: Molecular Biology Protocols: Carbohydrate Protocols

Pierce Protein Glycosylation Kits and Protocols. Pierce - [Read Pierce Protein Glycosylation Kits]

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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