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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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Fluorescent Staining of Cell Surface Antigens on Viable Cells Using Direct Immunofluorescence - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4439

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Immunofluorescence Staining Protocols

Protocol provides a method for analysis of cell surface antigens using direct immunofluorescence, i.e., when the antibody being used is directly labeled to a fluorescent dye. After staining, the cells can be analyzed immediately by flow cytometry or fixed and stored for later analysis. - [Read Fluorescent Staining of Cell Surface Antigens on Viable Cells Using Direct Immunofluorescence]

Glass Antibody Chip Protocol Nunc - http://www.biokom.com.pl/nunc/news/pdf/Nunc%20MicroArray%20Slides%20-%20glass%20protein%20binding.pdf

Category: Antibody Microarray Protocols

Glass Antibody Chip Microarray Protocol Nunc - [Read Glass Antibody Chip Protocol Nunc]

High Throughput Western Blotting Protocol for Antibody Quality Control - http://discover.nci.nih.gov/abminer/files/Western.pdf

Category: Western Blot Protocols

Protocol for high throughput western blotting for antibody quality control. Includes: SDS-PAGE; TRANSFER; IMMUNOSTAINING; DETECTION. - [Read High Throughput Western Blotting Protocol for Antibody Quality Control]

Human T Cell Activation Protocol - http://www.ebioscience.com/ebioscience/appls/AC145.htm#human

Category: Immunology: T Cell Protocols

Protocol for the Stimulation of human peripheral blood mononuclear cells with anti-human CD3 monoclonal antibody; MTT assay for detection of cellular proliferation. Human PBMCs can be activated in vitro by soluble anti-human CD3 antibodies. Performed titration studies with these antibodies and established the following protocol for stimulation of PBMC. - [Read Human T Cell Activation Protocol]

Hybridization and Detection Using the HC ExpressArray Kit Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4122

Category: Microarray Protocols: Microarray Hybridization Protocols

Protocol describes the direct detection of RNA on DNA microarrays using Hybrid Capture (HC) technology and the HC ExpressArray Kit developed by Diagene. The kit uses a proprietary antibody that binds specifically to RNA:DNA hybrids and a second, fluorescently labeled, antibody that detects the primary antibody. Total RNA is applied directly to a glass-spotted DNA microarray, and stable RNA:DNA hybrids are visualized via a Cy3-labeled secondary antibody. - [Read Hybridization and Detection Using the HC ExpressArray Kit Protocol]

Immunoaffinity Purification: Eluting Antigens from Immunoaffinity Columns Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4276

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

This elution procedure relies on disrupting the bonds between the antibody and the antigen. - [Read Immunoaffinity Purification: Eluting Antigens from Immunoaffinity Columns Protocol]

Immunoblot Protocol - http://cgap-mf.nih.gov/Protocols/DNARNAProteomicAnalysis/Proteomics/Immunoblot.html

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Protocol for Immunoblot. Includes: Staining and Laser Capture Microdissection; Protein Separation by Polyacrylamide Gel Electrophoresis; Electrophoretic Transfer To a Membrane (Nylon, PVDF or Nitrocellulose); Primary and Secondary Antibody Incubations; Visualization. - [Read Immunoblot Protocol]

Immunoblotting: Antigen Detection Using Chemiluminescence Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4271

Category: Western Blot Protocols: Antigen Detection Methods Protocols

The blot is blocked to prevent nonspecific adsorption of the immunological reagents. Antibodies are then bound to the proteins immobilized on the membrane, and the antigen is detected by labeling the antibodies with conveniently identified tags. Common labeling methods for chemiluminescent detection include anti-immunoglobulin antibody-coupled enzymes such as horseradish peroxidase, which catalyzes the oxidation of luminol and in turn releases light. - [Read Immunoblotting: Antigen Detection Using Chemiluminescence Protocol]

Immunoblotting: Preparing Immunoprecipitated Proteins Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4300

Category: Western Blot Protocols

Protein immunoprecipitation can be a useful preparative step for immunoblotting. For very rare proteins, the protein of interest can be purified and concentrated by standard immunoprecipitation techniques before immunoblotting. In addition, protein-protein interactions can be tested with an immunoprecipitating antibody that is specific for one protein of a complex and an immunoblotting antibody that is specific for a second member of the complex. - [Read Immunoblotting: Preparing Immunoprecipitated Proteins Protocol]

Immunocytochemistry - http://cancer.ucsd.edu/howelllab/Immunocyto.html

Category: Immunology: Immunocytochemistry Protocols

Method for plating cells, fixing cells, antibody incubation and mounting of slides. PHEM buffer, Antifade recipes. Howell Lab. - [Read Immunocytochemistry]

Immunocytochemistry in Free-Floating Sections Protocol - http://www.fisher.co.uk/techzone/life/table-pdfs/protocol/immunocytochemistry_netwells_protocol.pdf

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

There are several strategies to visualize the antibody. For transmitted light microscopy, color development substrates for enzymes are often used. The antibody can be directly labeled with the enzyme. However, such a covalent link between an antibody and an enzyme might result in a loss of both enzyme and antibody activity. For these reasons several multistep staining procedures have been developed, where intermediate link antibodies are used. In this protocol use the Vectastain ABC-kit. - [Read Immunocytochemistry in Free-Floating Sections Protocol]

Immunofluorescence Detection of RNAi-Induced Protein Knockdown in Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4344

Category: Immunology: Immunofluorescence Protocols

This protocol describes the use of a specific antibody that recognizes the targeted gene product to detect RNAi-induced gene knockdown in mammalian cells. Western blot technology can be used as an alternative (see Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting). - [Read Immunofluorescence Detection of RNAi-Induced Protein Knockdown in Mammalian Cells Protocol]

Immunofluorescence Detection of RNAi-Induced Protein Knockdown in Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4344

Category: RNAi Technology Protocols: RNAi Mammalian Cells Protocols

Protocol describes the use of a specific antibody that recognizes the targeted gene product to detect RNAi-induced gene knockdown in mammalian cells. Western blot technology can be used as an alternative. - [Read Immunofluorescence Detection of RNAi-Induced Protein Knockdown in Mammalian Cells Protocol]

Immunofluorescence Labeling of Cells Protocol - http://www.sigmaaldrich.com/Area_of_Interest/Life_Science/Antibody_Explorer/Procedures/Immunofluorescence.html

Category: Immunology: Immunofluorescence Protocols

Protocol for immunofluorescence labeling of cells. Includes: Cell Preparation; Fixation; Application of Primary Antibody; Application of Secondary Antibody and Evaulation. - [Read Immunofluorescence Labeling of Cells Protocol]

Immunofluorescence Staining Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E662E4DA09244177634DA520E076CB6&objectid=6673C6FEF0EF1F391890AEA1A3AE9FBD

Category: Immunology: Immunofluorescence Protocols: Immunofluorescence Staining Protocols

Provides a protocol for indirect immunofluorescence, which is a method that provides information about the locations of specific molecules and the structure of the cell. Antibody molecules for a specific target molecule are exposed to the cell or tissue being investigated. The binding of these molecules is detected by incubating the sample with a secondary antibody specific for immunoglobulin molecules and conjugated to fluorophore. - [Read Immunofluorescence Staining Protocol]

Immunofluorescence using Anti-Serotonin Antibody in C. elegans Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p1561

Category: C. elegans Protocols

Protocol for immunofluorescence using anti-serotonin antibody in C. elegans. - [Read Immunofluorescence using Anti-Serotonin Antibody in C. elegans Protocol]

Immunohistochemistry using Anti-Ganglioside Antibodies - http://www.glycotech.com/protocols/Proto2.html

Category: Molecular Biology Protocols: Carbohydrate Protocols

Indirect method measuring immunofluorescence coupled to second antibody. Best for membrane antigens in addition to intra- and extracellular antigens, may be applied to frozen tissue sections, to cells in suspension, and to cells attached to glass slides or coverslips. Tadashi Tai~Head, Department of Tumor Immunology, The Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan - [Read Immunohistochemistry using Anti-Ganglioside Antibodies]

Immunohistochemistry with AP-Conjugated (NBT/BCIP) Protocol - http://icg.cpmc.columbia.edu/cattoretti/Protocol/immunohistochemistry/NBTBCIPstaining.html

Category: Immunohistochemistry Protocols

Protocol for immunohistochemistry with AP-Conjugated (NBT/BCIP). Protocol extensively blocks slides, further diluting the primary antibody, lengthening the incubation and washing time, using a simple AP-conjugated secondary at high dilution and use a slow long development with the most powerful IHC development, NBT/BCIP. Includes: Single AP stainiing and Double AP staining. - [Read Immunohistochemistry with AP-Conjugated (NBT/BCIP) Protocol]

Immunoprecipitation of mRNA-Protein Complexes Protocol - http://www.nature.com/nprot/journal/v1/n2/pdf/nprot.2006.82.pdf

Category: Immunology: Immunoprecipitation Protocols

Protocol for immunoprecipitation of mRNA-protein complexes. In this protocol, an antibody targeting an RBP of interest is used to immunoprecipitate the RBP and any interacting molecules from a cell lysate. Reverse transcription followed by PCR is then used to identify individual mRNAs isolated with the RBP. This method focuses on examining an association between a specific RBP-mRNA complex, and it is best suited for a small scale screening of known or putative binding partners. - [Read Immunoprecipitation of mRNA-Protein Complexes Protocol]

Immunoprecipitation with Antibody:Agarose Conjugates Protocol - http://www.bdbiosciences.com/pharmingen/protocols/Agarose_Conjugates.shtml

Category: Immunology: Immunoprecipitation Protocols

Protocol for immunoprecipitation with antibody:agarose conjugates. Includes: Denaturing Conditions; Native Conditions; Pre-clearing. - [Read Immunoprecipitation with Antibody:Agarose Conjugates Protocol]

Immunoprecipitation: Denaturing Lysis Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4534

Category: Immunology: Immunoprecipitation Protocols

For many sources of antigens, one useful method of lysis is to treat cells with harsh, denaturing solutions to release most of the protein antigens, as described here. The lysates are then diluted to reduce the denaturing conditions to levels that are suitable for the formation of antibody-antigen complexes. The resulting solution is precleared prior to immunoprecipitation. - [Read Immunoprecipitation: Denaturing Lysis Protocol]

Immunoprecipitation: Preclearing the Lysate Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4535

Category: Immunology: Immunoprecipitation Protocols: Protein Immunoprecipitation Protocols

To reduce backgrounds and to improve the signal-to-noise ratio, an antibody that does not recognize the antigen being studied can be added to the lysate and processed as for a normal immunoprecipitation. Any nonspecific proteins that might contaminate the final immunoprecipitation step are presumably removed with this irrelevant antibody. - [Read Immunoprecipitation: Preclearing the Lysate Protocol]

Immunoscreening of Lambda-GT11 Bacteriophage Libraries with Antibody Probes - http://omrf.ouhsc.edu/~frank/GT11.html

Category: RNA Protocols: cDNA Libraries Library

Adsroption of Anti-E. coli Antibodies, cDNA Library Screening, Frank Lab. Oklahoma Medical Research Foundation - [Read Immunoscreening of Lambda-GT11 Bacteriophage Libraries with Antibody Probes]

Immunostaining Conditions - http://www.bcm.edu/rosenlab/protocols/IHC_conditions.pdf

Category: Immunohistochemistry Protocols

Immunostaining conditions. Includes: Antibody, Host, Source, Cat #, Antigen Retrieval, Block, Diluent, Ab Dilution and 2° antibody. - [Read Immunostaining Conditions]

In Situ Hybridization for Epstein Barr Virus-Encoded RNA (EBER) Protocol - http://icg.cpmc.columbia.edu/cattoretti/Protocol/immunohistochemistry/EBER-ISH4doublestain.html

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

Protocol for in situ hybridization for Epstein Barr virus-encoded RNA (EBER). Includes: Glassware decontamination; Double staining for EBER and a second antibody . - [Read In Situ Hybridization for Epstein Barr Virus-Encoded RNA (EBER) 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.