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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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Blocking Solutions Protocols - http://icg.cpmc.columbia.edu/cattoretti/Protocol/main_page_reagents.html#Blocking%20solutions

Category: Immunohistochemistry Protocols: Blocking Protocols

Protocols for blocking solutions. Includes: Endogenous peroxidase blocking solution (H2O2-Azide); Non-specific antibody background blocking solution: swine serum; Non-specific antibody background blocking solution: MILK; Enzymatic antigen retrieval solution; Unconjugated, biotin- and fluorochrome-conjugated antibody (primary) solutions; Unconjugated, biotin- and fluorochrome-conjugated antibody (secondary) solutions; Enzyme-conjugated secondary antibody solutions.; etc.. - [Read Blocking Solutions Protocols]

CD90 (Thy-1) Antibody Staining Protocol for Immunohistochemistry - http://www.ihcworld.com/_protocols/antibody_protocols/cd90_pharmingen.htm

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

Protocol for CD90 (Thy-1) antibody staining for Immunohistochemistry. Includes: Primary Antibody; Antigen Retrieval; Detection Methods; Chromogen Substrate; Counterstain. - [Read CD90 (Thy-1) Antibody Staining Protocol for Immunohistochemistry]

Cell Staining for Immunofluorescence Microscopy - http://www.bdbiosciences.com/pharmingen/protocols/Immunofluorescence_Microscopy.shtml

Category: Molecular Imaging: Immunofluorescence Microscopy

Cell Staining for Immunofluorescence Microscopy. Includes protocols for fixing the cells, Coverslip Preparation for Adherent Cells, Coverslip Preparation for Non-Adherent Cells, Paraformaldehyde Fixation, and Methanol/Acetone Fixation. Blocking protocols include blocking with primary antibody, and incubation with secondary antibody. - [Read Cell Staining for Immunofluorescence Microscopy]

Chemokine and Interleukin Antibody Protocols - http://www.capralogics.com/Protocols.htm

Category: Signalling Signal Transduction Protocols: Chemokine Protocols

Western Blotting chemokine receptors, Immunohistochemistry on Paraffin sections for chemokines. - [Read Chemokine and Interleukin Antibody Protocols]

Chromatin Immunoprecipitation (ChIP) of Protein Complexes Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4560

Category: Immunology: Immunoprecipitation Protocols: Chromatin Immunoprecipitation Protocols

Protocol describes the use of chromatin immunoprecipitation technology (ChIP) to analyze interactions of proteins or protein complexes with DNA in vivo. In this approach, the material is fixed with formaldehyde to preserve DNA-protein and protein-protein associations, the cells are lysed, and the chromatin is cut and solubilized. The chromatin suspension is immunoprecipitated with an antibody against the protein(s) of interest, and the coimmunoprecipitated DNA fragments are analyzed. - [Read Chromatin Immunoprecipitation (ChIP) of Protein Complexes Protocol]

Clontech Antibody Microarrays Protocol - http://www.bdbiosciences.ca/canada/downloads/Ab_Array_380_User_Manual.pdf

Category: Antibody Microarray Protocols

Clontech Antibody Microarrays Protocol. Clontech - [Read Clontech Antibody Microarrays Protocol]

Cytokine ELISA Protocol - http://www.bdbiosciences.com/pharmingen/protocols/Cytokine_ELISA.shtml

Category: Signalling Signal Transduction Protocols: Cytokine Protocols

Capture antibody, Blocking, Standards and Samples, Detection antibody, Cytokine ELISA Helpful Hints. BD Biosciences. - [Read Cytokine ELISA Protocol]

Cytokine Neutralization, In Vitro Using Antibodies - http://www.ebioscience.com/ebioscience/appls/NU.htm

Category: Signalling Signal Transduction Protocols: Cytokine Protocols

Antibody Neutralization of TNF-Î±-Induced Killing of L929 Cell Line, IFN-Î³-Protection from Viral Infection of L929 and A549 Cell Lines, TNF-Î±-Induced Killing of L929 Cell Line. eBiosciences. - [Read Cytokine Neutralization, In Vitro Using Antibodies]

Direct Elisa Using Primary Antibody Protocol - http://www.abcam.com/assets/pdf/protocols/Direct%20ELISA%20protocol.pdf

Category: ELISA Protocols

Direct Elisa Using Primary Antibody Protocol. Abcam. - [Read Direct Elisa Using Primary Antibody 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]

Early Events in B Lymphocyte Activation Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E6634190D367003ECF94AE65295FA77&objectid=6674AB8295693316922D8DFC029193A6

Category: Immunology: B Cell Protocols

B cell activation can be quantitated indirectly by assaying antibody production or directly by measuring cellular changes that occur immediately after exposure to an activation signal. Provides methods for the latter (direct) approach--namely, methods for quantifying early parameters of B cell activation such as increases in intracellular ionized calcium concentration [Ca2+]I, cell size, and MHC class II-antigen expression. - [Read Early Events in B Lymphocyte Activation Protocol]

Electron Microscopy of the Cytoskeleton of Cultured Cells - http://www.borisylab.northwestern.edu/pages/protocols/electmicrosctext.html

Category: Molecular Imaging: Electron Microscopy Protocols

The protocol for immunolabeling for electron microscopy depends mostly on the primary antibody and its ability to recognize antigen under particular ...Electron Microscopy of the Cytoskeleton of Cultured Cells. Boris Lab. - [Read Electron Microscopy of the Cytoskeleton of Cultured Cells]

ELISA for Antibody Isotype Determination Protocol - http://www.natureprotocols.com/2007/01/22/immunizations_and_enzymelinked.php

Category: ELISA Protocols

Protocol for immunizations and enzyme-linked immunosorbent assay (ELISA) for antibody isotype determination. - [Read ELISA for Antibody Isotype Determination Protocol]

ELISA Procedure for Measuring Serum Antibody Titer - http://www.komabiotech.com/technical/protocol/ELISA.htm

Category: ELISA Protocols

Method and ELISA Procedure for Measuring Serum Antibody Titer. Komabiotech - [Read ELISA Procedure for Measuring Serum Antibody Titer]

ELISPOT Protocol Detailed PDF - http://www.biolegend.com/images/Protocols/BioLegend%20ELISPOT%20protocol.pdf

Category: ELISA Protocols: ELISpot Protocols

Coat the Plate, Block the Plate, Add Detection Antibody, Solutions and buffers for ELISpot. Biolegend. - [Read ELISPOT Protocol Detailed PDF]

Enzyme-Linked ImmunoSorbent Assay (ELISA) for NGF Protocol - http://crutcherlab.med.uc.edu/ELI.htm

Category: Immunology: Immunoassays

Protocol for ELISA assay for NGF. Includes: ABSORPTION OF THE POLYCLONAL AND PREIMMUNE SERUM; BLOCKING; SAMPLE PREPARATION; PREPARATION OF NGF STANDARDS; PROTEIN RECOVERY; DESIGNING THE PLATE; APPLYING THE STANDARDS AND SAMPLES; APPLYING THE MONOCLONAL; APPLYING SECONDARY ANTIBODY; APPLYING STREPTAVIDIN; CHROMAGEN DEVELOPMENT; READING THE PLATE. - [Read Enzyme-Linked ImmunoSorbent Assay (ELISA) for NGF Protocol]

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]

Erythrophagocytosis Assay - http://www.icp.ucl.ac.be/mexp/vipg_eph.html

Category: Immunology: Macrophage Monocyte Protocols

Erythrophagocytosis Assay. The erythrophagocytosis assay is performed to compare the phagocytic rate with and without anti-eythrocyte antibody in either macrophages from control animals or virus-infected counterparts. Andrei Musaji Viral Immunity and Pathogenesis Group - [Read Erythrophagocytosis Assay]

FAK Autophosphorylation Assay - http://www.bio.com/protocolstools/protocol.jhtml?id=p1984

Category: Signalling Signal Transduction Protocols

This kinase assay is meant to determine whether an agonist or event can influence the autophosphorylation of FAK. The addition of 1 Î¼l of polyGT to the kinase reaction mix will determine the activity of the enzyme against a substrate. Includes information on: Harvest, Immunoprecipitation, Kinase Reaction and Antibody Detection of FAK. - [Read FAK Autophosphorylation Assay]

Fixation and Staining Parameters - http://www-biology.ucsd.edu/labs/aroian/protocols/fixatives.htm

Category: C. elegans Protocols: C. elegans Fixing Protocols

Parameters for fixation and staining. Includes: Antigen, Fixation, Fixation time, Antibody and Dilution. - [Read Fixation and Staining Parameters]

Fixing Caenorhabditis elegans in Bouin’s Fixative Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4521

Category: C. elegans Protocols: C. elegans Fixing Protocols

Bouin’s fixative is a particularly good choice for worms because it penetrates dense tissues well and is extremely good for fixing antigens. Like all strong fixatives, however, it is unsuitable for some antibody-antigen pairs. In such cases, the length of time in the Bouin’s fixative can be shortened, or paraformaldehyde fixation can be used instead. - [Read Fixing Caenorhabditis elegans in Bouin’s Fixative Protocol]

Flow cytometric determination of leukocyte surface antigens in whole blood - http://pingu.salk.edu/flow/protocols/wholeblood.html

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Flow cytometric determination of leukocyte surface antigens in whole blood. Quantitation of cell surface antigens in whole blood with the flow cytometer is very simple and requires:
1. Blood collection; 2. Addition of antibody; 3. Calibration of the flow cytometer; 4. Making measurements. - [Read Flow cytometric determination of leukocyte surface antigens in whole blood]

Flow Cytometric Determination of Leukocyte Surface Antigens in Whole Blood Protocol - http://pingu.salk.edu/flow/protocols/wholeblood.html

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

Protocol for flow cytometric determination of leukocyte surface antigens in whole blood. Includes: Blood collection; Add antibody to the blood; Make measurements. - [Read Flow Cytometric Determination of Leukocyte Surface Antigens in Whole Blood 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.