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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.]

A high-capacity RNA affinity column for the purification of human IRP1 and IRP2 overexpressed - http://www.rnajournal.org/cgi/reprint/9/3/364.pdf

Category: Chromatography Protocols: DNA RNA Affinity Chromatography

PDF. A high-capacity RNA affinity column for the purification of human IRP1 and IRP2 overexpressed in Pichia pastoris. CHARLES R. ALLERSON, ALAN MARTINEZ, EMINE YIKILMAZ and TRACEY A. ROUAULT. RNA Journal. - [Read A high-capacity RNA affinity column for the purification of human IRP1 and IRP2 overexpressed]

Affinity Antibody Purification of Protein A/G Chromatography - http://www.upstate.com/misc/protocol_detail.q.prot.e.antibody_purification.a.name.e.Antibody_Purification

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

Antibody Purification (Antiserum or Ascites by Protein A/G Chromatography). Species and Type of Antibody Agarose Rabbit IgG Protein A or Protein G Mouse IgG1 Protein G Mouse IgG2 Protein A or Protein G Mouse IgG3 Protein G Sheep IgG Protein G but binds weakly Rat IgG Protein G but binds weakly Guinea Pig IgG Protein A Dog IgG Protein A Goat IgG Protein G Pig IgG Protein A Hamster IgG Protein G. By Millipore. - [Read Affinity Antibody Purification of Protein A/G Chromatography]

Affinity Chromatography Handbook - http://books.google.com/books?id=kUl4k55F-fIC&pg=PA169&lpg=PA169&dq=dna+affinity+chromatography+protocol&source=web&ots=A7TBT29Ejf&sig=XkdH0cM4tNPKdN7y9XN2BfNMeno#PPA79,M1

Category: Chromatography Protocols

Affinity Chromatography Handbook. Google Books. - [Read Affinity Chromatography Handbook]

Affinity Chromotography Magnetic Bead Purification of RNA-binding proteins from Rat Brain. - http://147.52.72.117/IJMM/2003/volume11/number4/509.pdf

Category: RNA Protocols: RNA-binding Protein Purification

Affinity Chromotography Magnetic Bead Purification of RNA-binding proteins from Rat Brain. Scaturro et al. 2003. - [Read Affinity Chromotography Magnetic Bead Purification of RNA-binding proteins from Rat Brain.]

Affinity Purification of Antibodies - http://cmgm.stanford.edu/~vesicle/protocols/affinity.html

Category: Protein Protocols: Antibody Purification Protocols

Affinity Purification of Antibodies. Vesicle Trafficking. - [Read Affinity Purification of Antibodies]

Affinity purification of antibodies from crude serum - http://wheat.pw.usda.gov/~lazo/methods/au/sect32.html

Category: Protein Protocols: Antibody Purification Protocols

Affinity purification of antibodies from crude serum. David Bowtell Lab. - [Read Affinity purification of antibodies from crude serum]

Affinity purification of antibodies Protocol - http://www.xenbase.org/xmmr/Marker_pages/methods_pages/affinity_col.html

Category: Protein Protocols: Antibody Purification Protocols

Affinity purification of antibodies. Protocol by Peter Vize, XMMR - [Read Affinity purification of antibodies Protocol]

Affinity Purification of Interacting Proteins from Cell Lysates Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4593

Category: Chromatography Protocols: Affinity Chromatography Protocols

Recombinant protein or a chemically synthesized bioactive fragment is immobilized on resin and used as a probe to capture interacting proteins directly from a cell extract. Affinity-purified proteins are fractionated by gel electrophoresis and visualized by Coomassie staining. Proteins that interact specifically are identified by comparing this gel profile to one obtained from cell lysates passed over a control resin lacking the immobilized probe protein. - [Read Affinity Purification of Interacting Proteins from Cell Lysates Protocol]

Analysis of Oligosaccharide Ligands by High Performance Liquid Affinity Chromatography - http://www.glycotech.com/protocols/Proto6.html

Category: Molecular Biology Protocols: Carbohydrate Protocols

High performance liquid affinity chromatography (HPLAC) is a useful procedure to investigate he interactions between carbohydrate binding protein and their ligands. Technical requirements are similar to conventional HPLC. HPLAC can screen and separate natural ligands from complex biological mixtures. WeiTong Wang~GlycoTech Corporation, Rockville, Maryland - [Read Analysis of Oligosaccharide Ligands by High Performance Liquid Affinity Chromatography]

Analysis of Oligosaccharide Ligands by High Performance Liquid Affinity Chromatography - http://www.glycotech.com/protocols/Proto6.html

Category: HPLC Protocols

Analysis of Oligosaccharide Ligands by High Performance Liquid Affinity Chromatography Protocol. Glycotech. - [Read Analysis of Oligosaccharide Ligands by High Performance Liquid Affinity Chromatography]

Antibody Purification - http://axon.med.harvard.edu/~cepko/protocol/mike/A1.html

Category: Protein Protocols: Antibody Purification Protocols

This protocol includes an ammonium sulfate cut, affigel blue chromatography and affinity chromatography. Mike A. Dyer Lab Harvard. - [Read Antibody Purification]

Antibody Purification - Antiserum or Ascites by Protein A/G Chromatography - http://www.upstate.com/misc/protocol_detail.q.prot.e.antibody_purification.a.name.e.Antibody_Purification

Category: Protein Protocols: Antibody Purification Protocols

Antibody Purification (Antiserum or Ascites by Protein A/G Chromatography). Buffer Preparation, Preparation of a Protein A Agarose or Protein G Agarose Affinity Column, Pouring the Protein A/G Affinity Column, Preparation of Antiserum or Ascites for Affinity Chromatography, Affinity Chromatography Using Protein A/G Agarose. - [Read Antibody Purification - Antiserum or Ascites by Protein A/G Chromatography]

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]

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]

Construction of Micro-Tip for Use in IMAC Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000163.pdf?pid=PP00000163

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Protocol is the second step in a three-step process for the preparation and enrichment of phosphopeptides using immobilized metal affinity chromatography (IMAC) for the identification of the phosphopeptides by LCMS/ MS. This procedure describes the construction of microchromatographic columns, or micro-tips. - [Read Construction of Micro-Tip for Use in IMAC Protocol]

Coupling Peptides To Resin For Affinity Purification - Claire Walczak. - http://mitchison.med.harvard.edu/protocols/ab3.html

Category: Peptide Protocols: Antibodies from Peptides Protocols

Coupling Peptides To Resin For Affinity Purification - Claire Walczak. - [Read Coupling Peptides To Resin For Affinity Purification - Claire Walczak.]

Detection of DNA in Agarose Gels Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4022

Category: Nucleic Acid Electrophoresis Protocols: DNA Electrophoresis Protocols: DNA Electrophoresis in Agarose Gels Protocols

The most convenient and commonly used method to visualize DNA in agarose gels is staining with the fluorescent dye ethidium bromide. Ethidium bromide can be used to detect both singleand double-stranded nucleic acids (both DNA and RNA). However, the affinity of the dye for single-stranded nucleic acid is relatively low and the fluorescent yield is comparatively poor. - [Read Detection of DNA in Agarose Gels Protocol]

Detection of Fungi by Fluorescence Microscopy Using Fluorescent Brighteners - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./flbr.html~main

Category: Microscopy Protocols: Fluorescence Microscopy Protocols

Certain fluorescent dyes such as Blankophor have a high affinity for the b -glycosidically linked polysaccharides such as glucan and chitin, which are main the constituents of the fungal cell wall. Therefore, these fluorescent dyes can be used for screening clinical samples for the presence of fungal elements. This procedure can be performed using the following specimens: Nail, Skin, Bronchial alveolar lavage fluid, Sputum and Biopsies. - [Read Detection of Fungi by Fluorescence Microscopy Using Fluorescent Brighteners]

Detection of Ubiquitylated Proteins in Yeast Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4615

Category: Yeast Protocols: Yeast Protein Protocols

This assay is performed to detect ubiquitylated proteins in yeast. Yeast that have been transformed with a vector expressing polyhistidine-tagged ubiquitin (Ub) under the control of a copper-inducible promoter are grown, induced with copper, and harvested. Total ubiquitylated proteins are then recovered by nickel-affinity chromatography, and specific proteins are detected by Western blotting. - [Read Detection of Ubiquitylated Proteins in Yeast Protocol]

Direct Selection of cDNAs with Large Genomic DNA Clones Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4077

Category: Genetics and Genomics: Genotyping Protocols: Mutation Detection Protocols

The goal of this method is to identify transcriptionally active genes in cloned segments of genomic DNA. The protocol uses hybridization and affinity purification to recover biotin-labeled cDNAs that bind to a 500-kb segment of human DNA cloned in a BAC vector. However, the method can be easily adapted to other clones of genomic DNAs cloned in high-capacity vectors. - [Read Direct Selection of cDNAs with Large Genomic DNA Clones Protocol]

Directed Orientation of Antibodies during Preparation of Antibody Affinity Resin Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4204

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

Protocol describes the directed coupling of antibody to protein G-agarose via its Fc domain, and subsequent covalent cross-linking of the complex using dimethyl pimelimidate (DMP). - [Read Directed Orientation of Antibodies during Preparation of Antibody Affinity Resin Protocol]

DNA Affinity Chromatography Using Gravity Flow - Subscription Required - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4206

Category: Chromatography Protocols: DNA RNA Affinity Chromatography

DNA Affinity Chromatography, DNA affinity chromatography can be a low-tech method using gravity flow at 4Â°C, a disposable chromatography column, and DNA affinity resin prepared in the laboratory (see Preparation of a DNA Affinity Column). Include 10-20% glycerol and 0.025-0.1% NP-40 in the column buffers to suppress losses due to nonspecific adsorption of protein to surfaces. Load the protein in a buffer that is compatible with binding of the protein to its target site. Keith Brocklehurst et al - [Read DNA Affinity Chromatography Using Gravity Flow - Subscription Required]

DNA-Binding Protein Purification with Dynabeads. - http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=2671937

Category: DNA Protocols: EMSA DNA-Protein Protocols

Magnetic DNA affinity purification of yeast transcription factor tau--a new purification principle for the ultrarapid isolation of near homogeneous factor. Gabrielsen et al. 1989 - [Read DNA-Binding Protein Purification with Dynabeads.]

Articles (1-5 of 5)

Tubulin Polymerization Protocol using GTP and GMPCPP

Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.

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.

Antibodies Phage Expression Protocol Using 96-well Microtiter Plates

A Phage display expression protocol for antibody expression in 96-well microtiter plates.

Immobilized Antigen Phage Antibody Selection Protocol

A protocol for the selection of Phage Antibodies using Immobilized Antigen. This method describes the selection of antibodies from bacteriophage antibody libraries that recognize a specific antigen. The phage display library of antibody-displaying phage particles is exposed to antigen attached to a solid substrate (Nunc Immuno™ tubes). The phage particles with affinity for antigen bind to the immobilized antigen and are selected from the library of phage expressing antibodies.