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ANTIBODY PURIFICATION by affinity chromatography - http://mullinslab.ucsf.edu/protocols/html/ANTIBODY_PURIFICATION_by_affini.htm

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

ANTIBODY PURIFICATION by affinity chromatography. By Beth, Mullins Lab UCSF. To affinity purify antibodies, generate lots of E. coli lysate that contains your antigen. If the protein can stand freeze thawing, then go ahead and purify the protein from e. coli lysate and keep it frozen until you need to couple it to a CH-sepharose column. - [Read ANTIBODY PURIFICATION by affinity chromatography]

Chelating Sepharose Fast Flow Protocol - http://www.bio.indiana.edu/~ybelab/procedures/ChelatingSepharose.pdf

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Immobilized metal ion affinity chromatography (IMAC) exploits a molecule’s affinity for chelated metal ions. The amino acid histidine present in many proteins forms complexes with transition metal ions such as Cu2+, Zn2+, Ni2+ and Fe3+. Chelating Sepharose™ Fast Flow with a suitable immobilized metal ion will therefore selectively retain proteins with exposed histidine. - [Read Chelating Sepharose Fast Flow Protocol]

Construction of cDNA Libraries Stage 5: Fractionation of cDNA by Gel Filtration through Sepharose - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3730

Category: DNA Protocols: cDNA Library Protocols

Protocol for construction of cDNA library, stage 5: Fractionation of cDNA by Gel Filtration through Sepharose CL-4B. - [Read Construction of cDNA Libraries Stage 5: Fractionation of cDNA by Gel Filtration through Sepharose]

Kinase Assay Using In Vitro Translated Xenopus Mos Kinase - http://www.bio.com/protocolstools/protocol.jhtml?id=p912

Category: Signalling Signal Transduction Protocols

Protocol describes how tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay. Protocol includes: Translation of Xenopus Mos Kinase; Antibody to Antigen Binding; Protein A Sepharose to Antibody Binding; Kinase Reactions on Immunoprecipitated Material; Polyacrylamide Gel Analysis of Immunoprecipitates. Includes protocol hints. - [Read Kinase Assay Using In Vitro Translated Xenopus Mos Kinase]

Lectin-Agarose Affinity Chromatography Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4202

Category: Chromatography Protocols: Affinity Chromatography Protocols

Two methods are provided for purifying glycoproteins using wheat-germ agglutinin or concanavalin A-Sepharose. Because lectin-affinity matrices can bind a few milligrams of protein per milliliter of affinity matrix, only a small amount of affinity gel matrix is required. The batchwise method is recommended when protein volume is large. - [Read Lectin-Agarose Affinity Chromatography Protocol]

Preparation of DNA Affinity Resin - Subscription Required - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4205

Category: Chromatography Protocols

This protocol describes the preparation of concatamerized oligonucleotides and their coupling to cyanogen-bromide-activated Sepharose. The procedure uses a commercially activated resin, which can be purchased as a lyophilized powder. Keith Brocklehurst et al. - [Read Preparation of DNA Affinity Resin - Subscription Required]

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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.