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Analysis of Proteins by Immunoprecipitation - http://dps.ufl.edu/hansen/protocols/imp98.prt.htm

Category: Protein Protocols: Immunoprecipitation Protocols

Antibody-antigen complexes are removed from solution by addition of an insoluble form of an antibody binding protein such as Protein A, Protein G or second antibody. Immunoprecipitation protocols / methodology and technical background information. P.J. Ha - [Read Analysis of Proteins by Immunoprecipitation]

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]

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]

Large scale nuclear extract preparation - http://mesentsev.narod.ru/Biblioteque/Hattori.html

Category: Protein Protocols: Nuclear Extraction Protocols

Homogenize cells or tissue, Pellet nuclei, Lyse nuclei, Reprecipitate nuclear proteins, dialyze nuclear extract. Hattori M et al., DNA Cell Biol. 1990. Homogenization buffer and dialysis buffer recipes. - [Read Large scale nuclear extract preparation]

Analysis of Protein Gels (SDS-PAGE) - http://www.ruf.rice.edu/~bioslabs/studies/sds-page/gellab3.html

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)]

ChIP on ChIP Protocol PDF - http://genomecenter.ucdavis.edu/farnham/farnham/protocols/MOJO%20Dec%202003%20ChIP-chip%20protocol.pdf

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

Method to identify genomic targets of DNA-binding factors. Chromatin immunoprecipitation (ChIP) assay on high-throughput microarray based methods for discovering genomic regions occupied by human DNA-binding proteins. Oberley and Farnham. - [Read ChIP on ChIP Protocol PDF]

SDS-PAGE - http://www.ciwemb.edu/labs/koshland/Protocols/Protein/sdspage.html

Category: Protein Protocols: Protein Electrophoresis: SDS-PAGE Electrophoresis Protocols

SDS-PAGE: gel electrophoresis of proteins. Very simple protocol for SDS-PAGE. Koshland Lab, Carnegie Institution of Washington, Howard Hughes Medical Institute. - [Read SDS-PAGE]

Immunoprecipitation TroubleShooting Guide Sigma - http://www.sigmaaldrich.com/img/assets/8181/ImmunoprecipTroubleshoot.pdf

Category: Protein Protocols: Immunoprecipitation Protocols: Immunoprecipitation Troubleshooting

Immunoprecipitation TroubleShooting Guide. Sigma Aldrich. Problems: No binding, High background or unwanted proteins precipitate. With Solutions. - [Read Immunoprecipitation TroubleShooting Guide Sigma]

Column Chromatography Protocol - http://www.biochem.arizona.edu/classes/bioc463a/Info/protocols/exp_4_1.pdf

Category: Chromatography Protocols: Column Chromatography Protocols

Size Exclusion Column Chromatography Protocol. PDF. In this protocol you will learn how to use three types of column chromatography: Gel Filtration or Size Exclusion (SEC), Ion Exchange (IEC), and affinity (AC) in order to purify proteins and enzymes based on the physical properties of these biomolecules. Univ. Arizona, Biochemistry. - [Read Column Chromatography Protocol]

Troubleshoot Immunoprecipitation - http://www.chemicon.com/resource/ANT101/a2.asp#TroubleShooting

Category: Protein Protocols: Immunoprecipitation Protocols: Immunoprecipitation Troubleshooting

Troubleshoot Immunoprecipitation. Chemicon. The most common challenge with immunoprecipitation is trying to lower the number and type of background proteins that contaminate the washed immune complexes. Suggestions for decreasing background in IP. - [Read Troubleshoot Immunoprecipitation]

General Overview of Histology - http://www-bioc.rice.edu/bios576/histology/histology.htm

Category: Histology Protocols: Fixation of Cells Tissues Protocol

Fixation must convert the soluble contents of the cell (ie proteins, carbohydrates, ... http://www.protocol-online.org/prot/Histology/Fixation/ ...General Overview of Histology and Tissue Fixation. BIOS 576. Rice. - [Read General Overview of Histology]

Fractional Precipitation of Proteins by Ammonium Sulfate - Subscription - http://www.cshprotocols.org/cgi/content/full/2006/16/pdb.prot4309

Category: Protein Protocols: Protein Precipitation Procols

Fractional Precipitation of Proteins by Ammonium Sulfate Simpson Cold Spring Harbor Protocols.2006; 2006: pdb.prot4309 - [Read Fractional Precipitation of Proteins by Ammonium Sulfate - Subscription]

Cytokine ELISA Protocol - http://www.bdbiosciences.com/pharmingen/protocols/Cytokine_ELISA.shtml

Category: ELISA Protocols: Sandwich ELISA Assay Protocols

Cytokine sandwich ELISA are sensitive enzyme immunoassays that can specifically detect and quantitate the concentration of soluble cytokine and chemokine proteins. BD Biosciences - [Read Cytokine ELISA 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]

Troubleshooting the immunoprecipitation procedure IP and IB - Roche - http://www.roche-applied-science.com/PROD_INF/MANUALS/epitope/p66.pdf

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]

Detection of Recombinant Proteins by ELISA assay - http://omrf.ouhsc.edu/~frank/ELISA.html

Category: ELISA Protocols

Detection of Recombinant Proteins by ELISA protocol. Dr. Frank, Oklahoma Medical Research Foundation. - [Read Detection of Recombinant Proteins by ELISA assay]

Methanol-Chloroform Precipitation of Proteins - http://www.healthsystem.virginia.edu/internet/biomolec/methanolchloroform.cfm

Category: Mass Spectrometry Protocols

Methanol-Chloroform Precipitation of Proteins. A procedure for precipitating proteins from solution, including detergent solutions, is from Wessel and FlÃ¼gge (1984). Univ. Virginia. - [Read Methanol-Chloroform Precipitation of Proteins]

Extraction and analysis of diagnostically useful proteins from formalin-fixed, paraffin-embedded tis - http://www.jhc.org/cgi/content/full/46/3/397

Category: Protein Protocols: Protein Extraction From Tissue

Extraction and analysis of diagnostically useful proteins from formalin-fixed, paraffin-embedded tissue sections. Ikedaa et al, 1998 JHC. - [Read Extraction and analysis of diagnostically useful proteins from formalin-fixed, paraffin-embedded tis]

Transient Transfection Into 293T Cells Protocol - http://www.cbrinstitute.org/labs/springer/protocols/jun_293T.html

Category: Transfection Protocols: Transient Transfection Protocols

Transient transfection into 293T cells is a convenient way to overexpress and obtain both cellular and extracellular (secreted or membrane) proteins. 293 is a human renal epithelial cell line which is transformed by adenovirus E1A gene product. 293T is a derivative which also express SV40 large T antigen, allowing episomal replication of plasmids containing the SV40 origin and early promoter region. They (both) have the unusual property of being highly transfectable. - [Read Transient Transfection Into 293T Cells Protocol]

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]

A high yield affinity purification method for specific RNA-binding proteins. - http://www.pubmedcentral.gov/pagerender.fcgi?tool=pmcentrez&pageindex=1&artid=330202

Category: RNA Protocols: RNA-binding Protein Purification

A high yield affinity purification method for specific RNA-binding proteins: isolation of the iron regulatory factor from human placenta. Neupert 1990. - [Read A high yield affinity purification method for specific RNA-binding proteins.]

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]

Constructing and Expressing GFP Fusion Proteins - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4649

Category: Reporter Gene Assays: GFP Assay Protocols

The following protocol can be used for the development of stable cell lines expressing GFP fusion proteins. Although optimal transfection procedures (e.g., calcium phosphate, electroporation, or FuGENE 6 [Roche Applied Science]) vary depending on cell type, this general transfection procedure has been successful for stable transfection of HeLa, A-431, U2OS, BHK, and HT1080 cells. - [Read Constructing and Expressing GFP Fusion Proteins]

Cryosectioning Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/C7.html

Category: Immunohistochemistry Protocols: Cryosectioning Protocols

Protocol for cryosectioning. While the timing of the various steps in this protocol are probably not critical, process the tissue all at once to ensure that RNA and/or proteins do not get degraded. Includes: 20% Paraformaldehyde/4% Paraformaldehyde-PBS; Sucrose/PBS. - [Read Cryosectioning Protocol]

Simple and Reliable Method To Precipitate Proteins from Bacterial Culture Supernatant PDF - http://aem.asm.org/cgi/reprint/70/1/610.pdf

Category: Protein Protocols: Protein Precipitation Procols

Simple and Reliable Method To Precipitate Proteins from Bacterial Culture Supernatant. Randolph B. Caldwell et al., AEM ASM. - [Read Simple and Reliable Method To Precipitate Proteins from Bacterial Culture Supernatant PDF]

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.