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Preparation of Lysates Containing Fusion Proteins Encoded by Bacteriophage {lambda}: Lytic Infection - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3771

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

This rapid method is used to screen bacteriophage {lambda}gt11 clones for the production of immunodetectable fusion proteins. After optimizng the conditions of infection and induction, the method can be used to produce preparative amounts of a fusion protein. - [Read Preparation of Lysates Containing Fusion Proteins Encoded by Bacteriophage {lambda}: Lytic Infection]

Printing proteins as microarrays for high-throughput function determination. - http://cgr.harvard.edu/macbeath/protocols/protein_microarrays.html

Category: Protein Microarray Protocols

MacBeath G & Schreiber SL (2000). Printing proteins as microarrays for high-throughput function determination. Science 289: 1760-1763. MacBeath Lab - [Read Printing proteins as microarrays for high-throughput function determination.]

Protein Coimmunoprecipitation in Arabidopsis Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4683

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]

Protein Ligand Affinity Chromatography Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4203

Category: Chromatography Protocols: Affinity Chromatography Protocols

Protocol describes the coupling of peptides or proteins to Affi-Gel 10 (Bio-Rad). Affi-Gel 10 is a cross-linked agarose derivatized with N-hydroxysuccinimide (NHS) ester on a 10-atom-long spacer arm. The NHS ester allows spontaneous coupling of proteins via free amino groups. - [Read Protein Ligand Affinity Chromatography Protocol]

Protein Precipitation - http://www.aai.org/committees/education/Curriculum/protein.htm

Category: Protein Protocols: Protein Precipitation Procols

Protein Precipitation. Developed by: Eric Burtson. American Association of Immunologists 1995. In this lab you will separate a solution of proteins using protein precipitation. Since you will be testing the same protein mix that you used in the Gel Filtration lesson, you will pass the protein through a gel filtration column to identify which protein(s) precipitates. - [Read Protein Precipitation]

Protein Precipitation Microplate - http://www.htslabs.com/HTML/Wellplates/Applications/protein_precipitation_microplate.htm

Category: Protein Protocols: Protein Precipitation Procols

Laboratory sample cleanup is a necessary part for analytical preparation analysis. The removal of Contaminants such as proteins, cell debris and other materials is an important step. Typically this has been done by using Acetonitrile and then Centrifugation to pellet the debris leaving the clean supernant. After this process supernatant can be used for further analysis by HPLC, GC, MS and other analysis tandem methods. HTS Labs. - [Read Protein Precipitation Microplate]

Protein Separation on Ceramic Hydroxyapatite Using High-Performance Liquid Chromatography Protocol - http://cshprotocols.cshlp.org/cgi/content/extract/2006/9/pdb.prot4268

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol describes the separation of proteins using ceramic hydroxyapatite and HPLC. - [Read Protein Separation on Ceramic Hydroxyapatite Using High-Performance Liquid Chromatography Protocol]

Protocol for Antiphosphotyrosine Western Blot Analysis - http://www.bio.com/protocolstools/protocol.jhtml?id=p697

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]

Protocol for Immunopurification of Tyrosine Phosphorylated Proteins - http://www.bdbiosciences.com/pharmingen/protocols/Tyrosine_Phosphorylated_Proteins.shtml

Category: Signalling Signal Transduction Protocols

Protocol for immunopurification of tyrosine phosphorylated proteins. Protocol includes: Cell Lysis (Non-Denaturing and Denaturing Conditions) and Purification. - [Read Protocol for Immunopurification of Tyrosine Phosphorylated Proteins]

Protocol for Polymerase III In Vitro Transcription - http://www.bio.com/protocolstools/protocol.jhtml?id=p1140

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

Protocol describes an RNA Polymerase III (Pol III) transcription assay using an extract or proteins of choice. Pol III is the polymerase responsible for transcribing 5S RNA, tRNAs, and other small RNAs. Î±-Amanitin inhibits Pol II transcription in the assay. The newly-transcribed, radiolabeled RNA is visualized by autoradiography following Urea Polyacrylamide gel electrophoresis. - [Read Protocol for Polymerase III In Vitro Transcription]

Protocol for Protein Extraction Using Proteomics - http://www.biotech.unl.edu/proteomics/mercaptoethanol.html

Category: Protein Protocols: Protein Extraction Protocols

Protocol for Protein Extraction Using Proteomics. Extraction of proteins from plant cells that are rich in compounds that interfere with the 2-Dimensional electrophoretic separation methods such as salts, organic acids, phenolics, pigments, terpenes, among others. A common protocol used in our lab for extraction proteins from plant tissues consists in the homogenization of mortar-grounded material in liquid nitrogen with an extraction buffer. - [Read Protocol for Protein Extraction Using Proteomics]

Protocols for Insect Cell Culture - http://www.hyclone.com/library/insectprotocols.htm

Category: Cell Culture Protocols: Insect Cell Culture Protocols

Includes protocols: Direct Adaptation to HyQ© SFX-Insect™ Medium; Sequential Adaptation to HyQ© SFX-Insect™ Medium; Adaptation of High Five™ Cells to HyQ© SFX-Insect™; Production of Recombinant Protein Using the Baculovirus Expression Vector System; Culturing Insect Cells and Production of Baculovirus Expressed Proteins in 2.8 L Fernbach Systems. Production of Autographa californica Multiple Capsid Nuclear Polyhedrosis Virus (rAcMNPV) Stock - [Read Protocols for Insect Cell Culture]

Protocols for Mass Spectrometry Analysis Preparation - http://www.st-andrews.ac.uk/~bmsmspf/protocols.htm#manual

Category: Mass Spectrometry Protocols

Protocols for Mass Spectrometry Analysis Preparation. Preparing gel slices for PMF or MS/MS , mass spec data interpretation, Submitting proteins in solution, Interpretation of LC-MS/MS data. BMS MASS SPECTROMETRY AND PROTEOMICS FACILITY - [Read Protocols for Mass Spectrometry Analysis Preparation]

Purification by affinity chromatography of H1Â° RNA-binding - http://147.52.72.117/IJMM/2003/volume11/number4/509.pdf

Category: Chromatography Protocols

Purification by affinity chromatography of H1Â° RNA-binding proteins from rat brain. MARIA SCATURRO et al. - [Read Purification by affinity chromatography of H1Â° RNA-binding]

Purification of Antibodies on an Antigen Column Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4284

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

Immunoaffinity purification of antibodies is used to purify antigen-specific antibodies from a preparation of polyclonal antibodies. Such purification is commonly needed in the production of antipeptide antibodies, where it is used to concentrate the desired antibodies and separate them from those raised against carrier proteins. It is also used for the more general purpose of removing unwanted, nonspecific binding activity from polyclonal antibody preparations. - [Read Purification of Antibodies on an Antigen Column Protocol]

Purification of Fusion Proteins by Affinity Chromatography on Glutathione Agarose Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3655

Category: Chromatography Protocols: Affinity Chromatography Protocols

Recombinant proteins, constructed in pGEX vectors, are fused to glutathione S-transferase (GST) and can be purified to near homogeneity by affinity chromatography on glutathione-agarose. Bound GST-fusion proteins are readily displaced from the column by elution with buffers containing free glutathione. - [Read Purification of Fusion Proteins by Affinity Chromatography on Glutathione Agarose Protocol]

Purification of GST-fusion proteins E. coli. (pGEX vector) Protocol - http://mullinslab.ucsf.edu/protocols/html/GST_fusion_expression.htm

Category: E Coli Protocols: E coli Protein Protocols

Protocol for the purification of GST-fusion proteins E. coli. (pGEX vector). - [Read Purification of GST-fusion proteins E. coli. (pGEX vector) Protocol]

Purification of Histidine-tagged Proteins by Immobilized Ni2+ Absorption Chromatography Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4088

Category: Chromatography Protocols: Affinity Chromatography Protocols

Recombinant proteins engineered to have a polyhistidine tail at either the carboxyl or amino terminus can easily be purified in one step by affinity chromatography on a resin carrying chelated nickel ions. Chromatography can be carried out in column or batch formats. After unbound proteins are washed away, the target protein is eluted using imidazole, which typically preserves the antigenic and functional features of the protein. - [Read Purification of Histidine-tagged Proteins by Immobilized Ni2+ Absorption Chromatography Protocol]

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]

Purification of Maltose-binding Fusion Proteins by Affinity Chromatography on Amylose Resin Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4087

Category: Chromatography Protocols: Affinity Chromatography Protocols

Foreign proteins fused to maltose-binding protein can be readily purified to near homogeneity by affinity chromatography on resins containing cross-linked polysaccharides such as amylose. - [Read Purification of Maltose-binding Fusion Proteins by Affinity Chromatography on Amylose Resin Protocol]

Rapid and Reliable Preparation of Total Cell Lysates from Fission Yeast Protocol - http://www.natureprotocols.com/2006/07/21/rapid_and_reliable_preparation.php

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

Protocol for rapid and reliable preparation of total cell lysates from fission yeast. Method for successful and reproducible extraction of proteins from S. pombe cells. - [Read Rapid and Reliable Preparation of Total Cell Lysates from Fission Yeast Protocol]

Red Blood Cell Test System Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/37.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cell Specific Cytotoxicity Assays Protocols

An in vitro red blood cell assay is presented which allows the estimation of the irritation potential of tensides and tenside containing materials such as shampoos, shower gels, cleaning products, etc. The estimation is based on the fact that surfactants interact strongly with cellular membranes and proteins. Both effects are measured photometrically by use of the inherent native dye, oxyhemoglobin. - [Read Red Blood Cell Test System Protocol]

Removal of Cross-reactive Antibodies from Antiserum: Affinity Chromatography Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3340

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

Protocol describes a method for removing antibodies that react with bacterially encoded proteins by passing a crude preparation of immunoglobulins through a column containing immobilized bacterial proteins. - [Read Removal of Cross-reactive Antibodies from Antiserum: Affinity Chromatography Protocol]

Removal of Cross-reactive Antibodies from Antiserum: Incubation with E. coli Lysate - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3767

Category: Antibody Protocols: Antibody Production Protocols

This protocol describes how antibodies that react with bacterial-encoded proteins may be removed from polyclonal antisera by incubation with a bacterial lysate. - [Read Removal of Cross-reactive Antibodies from Antiserum: Incubation with E. coli Lysate]

Articles (1-3 of 3)

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.

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.