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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]

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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]

MAPK immunoprecipitation Protocol PDF - http://www.upstate.com/img/coa/17-192-18181.pdf

Category: Signalling Signal Transduction Protocols: MAPK Protocols

MAPK immunoprecipitation protocol and methods. Upstate. - [Read MAPK immunoprecipitation Protocol PDF]

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]

Native Chromatin Immunoprecipitation Protocol - http://www.abcam.com/index.html?pageconfig=resource&rid=30&pid=5

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

Native Chromatin Immunoprecipitation Protocol ABCAM - [Read Native Chromatin Immunoprecipitation 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]

Polyclonal Antibody Production Protocol - http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Immunology/PAb.html

Category: Antibody Protocols: Antibody Production Protocols

Protocol for polyclonal antibody production. Very useful for rapid and simple generation of antibodies for western blots, ELISA assays, and immunoprecipitation. Includes: Rabbit Immunization; Initial Preparation; Pre-bleed; Antigen Injection; Monitoring of Titer; Purification of Antibodies. - [Read Polyclonal Antibody Production Protocol]

Preparation of Worm Extracts by Sonication Protocol - http://www.genetics.wustl.edu/tslab/Protocols/Cyto_extract_sonicat_v2.htm

Category: C. elegans Protocols

Protocol was designed to rapidly generate small scale cytosolic extracts of C. elegans for Western or IP (has not been tested for RNA work). The protocol works well for between 50 to 5000 worms and has not been extensively tested on larger a scale, though it should work. Includes: Collection; Sonication; Clearing lysate; Immunoprecipitation. - [Read Preparation of Worm Extracts by Sonication Protocol]

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 Immunoprecipitation from a Yeast Cell Lysate Protocol - http://www.fhcrc.org/science/labs/gottschling/yeast/yeastip.html

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Immunoprecipitation Protocols

Protocol for protein immunoprecipitation from a Yeast cell lysate. - [Read Protein Immunoprecipitation from a Yeast Cell Lysate Protocol]

Protocol for Competitive Ligand Binding to Cortical Receptor using Crosslinking - http://www.bio.com/protocolstools/protocol.jhtml?id=p1254

Category: Signalling Signal Transduction Protocols

Protocol describes a competitive ligand binding assay for cortical neurotrophin receptors. Following binding in the presence of competitor, the bound radiolabeled ligand is cross-linked to the receptor. The cells are lysed and the ligand-receptor complexes are immunoprecipitated using a pan-trk (tyrosine kinase receptor) antibody. Protocol includes:Preparation of Cortical Tissue for Competitive Crosslinking, Competitive Binding, Crosslinking Ligand to Receptor, Lysis and Immunoprecipitation etc - [Read Protocol for Competitive Ligand Binding to Cortical Receptor using Crosslinking]

Protocol for Immunoprecipitation - http://www.fgsc.net/neurosporaprotocols/Immunoprecipitation%20final.pdf

Category: Protein Protocols: Immunoprecipitation Protocols

Protocol for immunoprecipitation. - [Read Protocol for Immunoprecipitation]

Protocol Immunoprecipitation - http://www.sigmaaldrich.com/Area_of_Interest/Life_Science/Antibody_Explorer/Procedures/Immunoprecipitation.html

Category: Immunology: Immunoprecipitation Protocols

Immunoprecopitation method, the protein from the cell or tissue homogenate is precipitated in an appropriate lysis buffer by means of an immune complex which includes the antigen (protein), primary antibody and Protein A-, G-, or L-agarose conjugate or a secondary antibody-agarose conjugate - [Read Protocol Immunoprecipitation]

Pulse-Chase Assay for Measuring Protein Stability in Yeast Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4641

Category: Yeast Protocols: Yeast Protein Protocols

This assay directly measures the stability of a specific protein in yeast. Following pulse-labeling of yeast cells, the target protein is recovered by denaturing immunoprecipitation. - [Read Pulse-Chase Assay for Measuring Protein Stability in Yeast Protocol]

Raf-1 Kinase Assay - http://www.bio.com/protocolstools/protocol.jhtml?id=p1980

Category: Signalling Signal Transduction Protocols: MAPK Protocols

This protocol uses mitogen-activated protein/ERK kinase (MEK) to activate the extracellular-signal-regulated (Erk) mitogen-activated protein (MAP) kinases upon agonist binding to receptors. Protocol includes information about: Harvest, Immunoprecipitation and Kinase Reaction. Also includes following solutions: Laemmli Sample Buffer (1X), ATP Mix, Kinase Buffer (10X), Lysis Buffer. Helpful protocol hints are also included. - [Read Raf-1 Kinase Assay]

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]

Troubleshooting Immunoprecipitation - http://www.roche-applied-science.com/PROD_INF/MANUALS/epitope/p66.pdf

Category: Protein Protocols: Immunoprecipitation Protocols

Troubleshooting Immunoprecipitation. Roche. - [Read Troubleshooting Immunoprecipitation]

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]

Troubleshooting Tips for Immunoprecipitation / Kinase Assays - http://www.upstate.com/misc/protocols.q.prot.e.tipsipkinase/Troubleshooting+Tips+for+Immunoprecipitation+%2F+Kinase+Assays

Category: Signalling Signal Transduction Protocols: Protein Kinase Protocols

Troubleshooting Tips for Immunoprecipitation / Kinase Assays. Upstate. - [Read Troubleshooting Tips for Immunoprecipitation / Kinase Assays]

Xenopus Chromatin immmunoprecipitation Protocol - http://spot.colorado.edu/~klym/Methods/XChIP.htm

Category: Xenopus Protocols: Xenopus Genetics Protocols

Protocol for Xenopus chromatin immunoprecipitation. - [Read Xenopus Chromatin immmunoprecipitation Protocol]

Yeast Chromatin Immunoprecipitation (ChIP) - http://www.fhcrc.org/science/labs/hahn/methods/mol_bio_meth/hahnlab_ChIP_method.html

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

Chromatin IP Method, ChIP buffers, Notes on ChIP Method, Quantative PCR using 32P dCTP. Hahn Lab. - [Read Yeast Chromatin Immunoprecipitation (ChIP)]

Yeast Chromatin Immunoprecipitation (ChIP) Assay Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4642

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Immunoprecipitation Protocols

Protocol describes a method for the detection of proteins bound to specific regions of chromatin in yeast. There are many variations of this assay. - [Read Yeast Chromatin Immunoprecipitation (ChIP) Assay Protocol]

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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.

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