epitope Protocols

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Epitope Mapping by Competition Assay Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4277

Category: Antibody Protocols: Epitope Mapping Antibody Protocols

The simplest way to determine whether two monoclonal antibodies bind to distinct sites on a protein antigen is to carry out a competition assay. The assay can be used with antibodies that bind both conformational and linear epitopes, and it is most useful in the analysis of monoclonal antibody specificity because polyclonal sera typically recognize multiple different epitopes. - [Read Epitope Mapping by Competition Assay Protocol]

Epitope Mapping Using Synthetic Biotin-Labeled Peptides Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4278

Category: Antibody Protocols: Epitope Mapping Antibody Protocols

A set of overlapping synthetic peptides is synthesized, each corresponding to a small segment of the linear sequence of a protein antigen and arrayed on a solid phase. The panel of solid-phase peptides is then probed with a test antibody, and bound antibody is detected using an enzyme-labeled secondary antibody. This method is very rapid and can be extraordinarily successful. - [Read Epitope Mapping Using Synthetic Biotin-Labeled Peptides Protocol]

Epitope Tagging of Recombinant Proteins Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E663998FCCB61070F32F1EAE0301F7F&objectid=6677C749FA75FD435618AC07256968A7

Category: Immunology: Immunoprecipitation Protocols

Protocols for detection and purification of proteins tagged with a particular epitope, the FLAG tag, although the same general approach can be applied to other epitope tags. The protocols employ the anti-FLAG M2 antibody to detect and purify FLAG-tagged proteins. The methods presented are immunoprecipitation of FLAG fusion proteins from cells using an anti-FLAG M2 affinity gel, detection of FLAG fusion proteins by western blotting, and purification of FLAG fusion proteins by anti-FLAG. - [Read Epitope Tagging of Recombinant Proteins Protocol]

Immunoprecipitation of Epitope-Tagged Proteins from Yeast Protocol - http://www.med.unc.edu/~hdohlman/TAP.html

Category: Immunology: Immunoprecipitation Protocols: Protein Immunoprecipitation Protocols

Protocol for immunoprecipitation of epitope-tagged proteins from yeast. - [Read Immunoprecipitation of Epitope-Tagged Proteins from Yeast 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]

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 the Western blot procedure Roche PDF - http://www.roche-applied-science.com/PROD_INF/MANUALS/epitope/p34-35.pdf

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

Troubleshooting the Western blot procedure Roche. PDF - [Read Troubleshooting the Western blot procedure Roche PDF]

Using FLAG Epitope-Tagged Proteins for Coimmunoprecipitation of Interacting Proteins Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/28/pdb.prot4557

Category: Immunology: Immunoprecipitation Protocols: Protein Immunoprecipitation Protocols

Protocol describes the use of FLAG-epitope-tagged proteins for both small-scale analysis and large-scale coimmunoprecipitation of interacting proteins. When examining protein interactions, it is sometimes possible to immunoprecipitate an endogenous protein X directly, without using an epitope tag, if antibodies are available. The advantage of examining interactions of endogenously expressed proteins is that these are likely to be physiological and less likely to be an artifact of overexpression. - [Read Using FLAG Epitope-Tagged Proteins for Coimmunoprecipitation of Interacting Proteins Protocol]