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Protein and Antibody Microarray Chips - An Introduction - http://www.molecularstation.com/protein-microarrays/

Protein Microarray Chips - An Introduction. Introduction, Types of Protein Chips, Attachment, Protein and Antibody Chip Production, Applications of Protein Chips, Detection methods, and Future Directions. Molecularstation. - [Read Protein and Antibody Microarray Chips - An Introduction]

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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]

Labeling Antibodies with Biotin Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4286

Category: Antibody Protocols: Antibody Labeling Protocols

This protocol describes the covalent coupling of antibodies to biotin. Biotin groups bind with extremely high affinity to streptavidin and avidin, both of which are available commercially coupled with enzymes, fluorescent dyes, or iodine. A biotinylated primary antibody, therefore, can be detected with any of a wide variety of different labels. The biotinylation reaction is simple and mild, and rarely inactivates the antibody. - [Read Labeling Antibodies with Biotin Protocol]

Labeling Monoclonal Antibodies by Biosynthesis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4288

Category: Antibody Protocols: Antibody Labeling Protocols

This method for tagging monoclonal antibodies involves growing hybridomas in the presence of radioactive amino acids. This protocol can be particularly useful when conventional labeling techniques cause the antibody to lose activity. The labeled antibodies that result are essentially identical to the unlabeled antibodies. - [Read Labeling Monoclonal Antibodies by Biosynthesis Protocol]

Making antibodies to synthetic peptides - http://medicine.ucsd.edu/hypertension/list_of_protocols/16_antibodies_to_peptides.htm

Category: Peptide Protocols: Antibodies from Peptides Protocols

Protocol for Antibody Creation from a Synthetic Peptide. Antibody from Peptides. Daniel O'Connor. - [Read Making antibodies to synthetic peptides]

Measuring Protein Concentration by Western Analysis Using Enhanced Chemiluminescence Detection - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4255

Category: Western Blot Protocols: Antigen Detection Methods Protocols

Protocol describes, samples containing the target protein are deposited onto a polyvinyldifluoride (PVDF) membrane using a vacuum manifold. The immobilized protein is exposed to an antibody specific for the target protein, followed by an antibody that reacts with species-specific determinants carried by the primary antibody and is conjugated to horseradish peroxidase (HRP). - [Read Measuring Protein Concentration by Western Analysis Using Enhanced Chemiluminescence Detection]

Monoclonal Antibody Production - http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Immunology/MAb.html

Category: Antibody Protocols: Antibody Production Protocols

Monoclonal Antibody Production. Includes: Mouse Immunization; Bleeding Mice; Hybridoma Fusion; Assaying for Positive Clones; Expanding Positive Clones. - [Read Monoclonal Antibody Production]

Monoclonal Antibody Production Fusion Protocol - http://www.icp.ucl.ac.be/mexp/vipg_fus.html

Category: Antibody Protocols: Antibody Production Protocols

Protocol for monoclonal antibody production fusion. Fusion is an important procedure to produce monoclonal antibodies (MoAb). - [Read Monoclonal Antibody Production Fusion Protocol]

Monoclonal Antibody Screening by Direct ELISA Protocol - http://www.its.caltech.edu/~bjorker/ELISA_.pdf

Category: ELISA Protocols

Protocol for monoclonal antibody screening by direct ELISA. - [Read Monoclonal Antibody Screening by Direct ELISA Protocol]

MTT Assay for Detection of Cellular Proliferation - http://www.ebioscience.com/ebioscience/appls/AC145.htm#mouse

Category: Immunology: T Cell Protocols

Mouse Protocol: Stimulation of mouse peripheral T cells with plate-bound 145-2C11 monoclonal antibody; MTT assay for detection of cellular proliferation. - [Read MTT Assay for Detection of Cellular Proliferation]

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]

Phycoerythrin Conjugation Protocol - http://pingu.salk.edu/flow/protocols/pelabel.html

Category: Immunology

Protocol for phycoerythrin conjugation. Includes: Preparation of PE; SPDP modification of PE; SMCC modification of antibody; DTT treatment of SPDP-PE; Purification of reactants; Conjugation; Stop reaction; Concentrate product; Separate conjugate. - [Read Phycoerythrin Conjugation 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]

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 Immunogold Electron Microscopy - http://www.hopkinsmedicine.org/cellbio/devreotes/antibody.htm

Category: Microscopy Protocols: Electron Microscopy Protocols

Immunogold electron microscopy protocols for CHIFF labeling and whole cells. - [Read Protocol for Immunogold Electron Microscopy]

Protocol for Motility using VE-DIC Microscopy - http://www.proweb.org/kinesin/Methods/VE-DIC.html

Category: Cell Biology and Cell Culture

Protocol for motility using VE-DIC microscopy. Materials required: Anti-GST (glutathione S-transferase) antibody, PB buffer =10 mM NaPO4 pH 7.2, EGTA, MgCl2, Clarified motor lysate, MTs or Axoneme-MTs,MgÂ·ATP, VALAP (1 Vasoline: 1 Lanolin: 1 Paraffin, heated gently until melted) and Maxell XR-S120 Black Magnetite Super-VHS tapes or comparable. - [Read Protocol for Motility using VE-DIC Microscopy]

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]

Purification Of Anti Peptide Antibody - Claire Walczak - http://mitchison.med.harvard.edu/protocols/ab4.html

Category: Peptide Protocols: Antibodies from Peptides Protocols

Purification Of Anti Peptide Antibody - Claire Walczak - [Read Purification Of Anti Peptide Antibody - Claire Walczak]

Purification of Antibodies on an Antigen Column Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4284

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

Immunoaffinity purification of antibodies is used to purify antigen-specific antibodies from a preparation of polyclonal antibodies. Such purification is commonly needed in the production of antipeptide antibodies, where it is used to concentrate the desired antibodies and separate them from those raised against carrier proteins. It is also used for the more general purpose of removing unwanted, nonspecific binding activity from polyclonal antibody preparations. - [Read Purification of Antibodies on an Antigen Column Protocol]

RNA Whole Mount In Situ Hybridization Protocol - http://genetics.med.harvard.edu/~cepko/protocol/ctlab/ish.ct.htm

Category: Cytogenetics Protocols: In Situ Hybridization Tissue Sections Protocols

Protocol for RNA whole mount in situ hybridization. Includes: Embryo preparation; Prehybridization and Hybridization; Post-hybridization washes, blocking, and antibody incubation; Post-antibody washes; Color development. - [Read RNA Whole Mount In Situ Hybridization Protocol]

RUVKUN Antibody Staining Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=11

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

Protocol for RUVKUN antibody staining. Includes: Fixation; To make solution; REDUCING DISULFIDES TO -SH; OXIDIZE -SH GROUPS TO -SO3; TO CHECK THAT WORMS ARE PERMEABLE TO MACROMOLECULES; ANTIBODY INCUBATIONS; PERMANENT SPRINGTIME MOUNTING. - [Read RUVKUN Antibody Staining Protocol]

RUVKUN Antibody Staining Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=11

Category: C. elegans Protocols: C. elegans Staining Protocols

Protocol for RUVKUN Antibody Staining. Includes: Fixation; To make solution; REDUCING DISULFIDES TO -SH; OXIDIZE -SH GROUPS TO -SO3; TO CHECK THAT WORMS ARE PERMEABLE TO MACROMOLECULES; ANTIBODY INCUBATIONS; PERMANENT SPRINGTIME MOUNTING. - [Read RUVKUN Antibody Staining Protocol]

SAPK / Jun kinase assays - http://kinase.uhnres.utoronto.ca/methods/SAPKassays.html

Category: Signalling Signal Transduction Protocols

SAPK/Jun kinase assays. Preparation of cell lysate, Substrate/Antibody, Lysate Interaction and buffer recipes. Woodgett Lab PMH. - [Read SAPK / Jun kinase assays]

SAPK/Jun Kinase Assays - http://kinase.uhnres.utoronto.ca/methods/SAPKassays.html

Category: Signalling Signal Transduction Protocols: MAPK Protocols

Protocol for SAPK/Jun kinase assays. Protocol includes information on: preparation of cell lysate, substrate/antibody, lysate interaction. - [Read SAPK/Jun Kinase Assays]

Semi-Quantitative Measurement of Proteins by Dot Blotting - http://www.cbrinstitute.org/labs/springer/protocols/jun_dotblot.html

Category: Protein Protocols: Protein Quantitation Concentration Protocols

Semi-Quantitative Measurement of Proteins by Dot Blotting. Jun Takagi Springer Lab. Includes protein quantitation, nitrocellulose, antibody, chemiluminescence, peroxidase.

- [Read Semi-Quantitative Measurement of Proteins by Dot Blotting]

Simple Western Blot Protocol - http://www.prosci-inc.com/Antibody/Western-Blot-Procedure.html

Category: Protein Protocols: Western Blot Protocol

Simple Western Blotting Protocol ProSci. - [Read Simple Western Blot Protocol]

Articles (1-4 of 4)

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.