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A Sensitive ELISA-Based Assay for Glycosyltransferases - http://www.glycotech.com/protocols/Proto4.html

Category: Molecular Biology Protocols: Carbohydrate Protocols

The method allows the detection and quantification of glycosyltransferase activity using an ELISA-based procedure and carbohydrate-specific monoclonal antibodies. Avoids the use of radiolabeled substrates. Bruce A. Macher~Professor of Chemistry and Biochemistry, San Francisco State University, San Francisco, CA - [Read A Sensitive ELISA-Based Assay for Glycosyltransferases]

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]

Anti-Nogo-A Treatment Protocol - http://www.natureprotocols.com/2006/07/21/antinogoa_treatment_enhances_c.php

Category: Neuroscience Protocols

After section of the corticospinal (CS) tract in adult rats, neutralizing Nogo-A with monoclonal antibodies leads to enhancement of axonal re-growth and compensatory sprouting, accompanied by increased motor recovery. The neutralization of Nogo-A represents a promising approach for therapy after lesion if its enhancement of functional recovery can be transposed to primates. - [Read Anti-Nogo-A Treatment Protocol]

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]

Assays for B Lymphocyte Function Protocols - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E66340AD6485AA0FEB8F538B2FD389E&objectid=6674A538AC9B642C5CD51556EF7B3D26

Category: Immunology: B Cell Protocols

Describes the antigenic stimulation of in vitro antibody production by B cells and the subsequent measurement of secreted antibodies. The first protocol is a generalized system for inducing in vitro antibody production and can accommodate various types of antigens under study. Secreted antibodies can then be measured with an enzyme-linked immunosorbent assay (ELISA) or other soluble-antibody detection systems. - [Read Assays for B Lymphocyte Function Protocols]

Band Shift Protocols - http://www.pharma.ethz.ch/institute_groups/biomacromolecules/protocols/bandshift

Category: DNA Protein Interactions Protocols: Electrophoretic Mobility Shift Assays EMSA Protocols

Protocols on band shift. Includes: Phosphorylation of recombinant antibodies; Labeling the antigen with a red fluorophore; Native gel electrophoresis; Band-shift assay for the determination of Kd; Band-shift assay for the determination of koff. - [Read Band Shift Protocols]

Binding Antibodies to Tissue Sections Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4331

Category: Antibody Protocols: Antibody Labeling Protocols

Once tissues are fixed and permeabilized, the antibodies are added. These antibodies can be labeled directly or detected by a labeled secondary reagent. For indirect detection, any reagent that binds specifically to the primary antibody can be "tagged" and used to locate the antibody. The possible reagents include anti-immunoglobulin antibodies, protein A or G, or, if the first antibody is labeled with biotin, streptavidin. They can be labeled with enzymes or gold. - [Read Binding Antibodies to Tissue Sections Protocol]

Characterization of Cells by Flow Cytometry Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000018.pdf?pid=PP00000018

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol uses specific antibodies coupled to one of four fluorochromes: fluorescein isothiocyanate (FITC), R-phycoerythrin (PE), peridinin chlorophyll-a (PcP), and allophycocyanin (APC). These fluorochromes can be used simultaneously to stain and analyze the expression patterns of four different proteins in the same sample. The fluorochrome stained cell populations are analyzed using a FACSCalibur dual-laser flow cytometer. - [Read Characterization of Cells by Flow Cytometry Protocol]

Chromatin IP (CHIP assay) - http://www.fhcrc.org/science/labs/breeden/Methods/chromatinIP.html

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

ChIP assay protocol with 2 steps: in vivo formaldehyde cross-linking of whole cells and protein-protein and protein-DNA interactions, followed by immunoprecipitation of protein-DNA complexes with specific antibodies from sonicated extracts. Breeden Lab - [Read Chromatin IP (CHIP assay)]

Color Changing Karyotyping (CCK) Protocol: An M-FISH/SKY Alternative - http://info.med.yale.edu/genetics/ward/tavi/CCKmethod.html

Category: Cytogenetics Protocols

Protocol for color changing karytoping (CCK): an M-FISH/SKY alternative. Includes: Slide preparation and hybridization; Probes, dyes and antibodies; Probe detection and image capture; Image processing. - [Read Color Changing Karyotyping (CCK) Protocol: An M-FISH/SKY Alternative]

Confocal Microscopy Specimen Preparation Using Synthetic Fluorophores and Immunofluorescence - http://www.olympusconfocal.com/applications/protocols/index.html

Category: Microscopy Protocols: Confocal Microscopy Protocols

Includes staining protocols: Triple-Staining Adherent Cells with MitoTracker, Phalloidin,& Nuclear Dyes; Staining Adherent Cells with Cytokeratin Primary Antibodies & Synthetic Fluorophores; Staining Adherent Cells with Intermediate Filament Primary Antibodies & Synthetic Fluorophores; Staining Cells & Tissue Cryosections with Tubulin Primary Antibodies, Phallotoxins,& Synthetic Fluorophores; Triple-Staining Tissue Cryosections with Wheat Germ Agglutinin, Phalloidin,& Nuclear Dyes; etc. - [Read Confocal Microscopy Specimen Preparation Using Synthetic Fluorophores and Immunofluorescence]

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]

Detection FISH Protocol - http://www.riedlab.nci.nih.gov/publications/FISH%20Detection.pdf

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Protocol for Detection (FISH). TRITC, or Avidin Cy-5. For the probes labeled with digoxigenin, we usually first incubate with mouse-anti-digoxigenin, followed by incubation with sheep anti-mouse Cy5.5, or other fluorochrome conjugated antibodies. - [Read Detection FISH Protocol]

Detection of Autoantibodies with Self-Assembling Radiolabeled Antigen Tetramers Protocol - http://www.natureprotocols.com/2007/01/29/detection_of_autoantibodies_wi.php

Category: Immunology

Protocol for detection of autoantibodies with self-assembling radiolabeled antigen tetramers. Details how to produce radiolabeled antigen-streptavidin tetramers for detection of antibodies by immunoprecipitation. Optionally, the antigen tetramers can be denatured to compare responses to folded and unfolded antigen in the same system. This technique can be applied to a large or small number of samples, and a given sample can be simultaneously assayed with multiple antigens. - [Read Detection of Autoantibodies with Self-Assembling Radiolabeled Antigen Tetramers 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]

ELISA for detection of platelet-associated Ig (PAIg) - http://www.icp.ucl.ac.be/mexp/vipg_plelisa.html

Category: ELISA Protocols

Qualitative ELISA (Enzyme-Linked Immunosorbent Assay) used for screening detection of anti-platelet antibodies or for detection of platelet-associated Ig (PAIg). Andrei Musaji Viral immunity and pathogenesis group, Universite Catholique de Louvain. - [Read ELISA for detection of platelet-associated Ig (PAIg)]

ELISA for Detection of Xenopus Antibodies Against FV3 Protocol - http://www.urmc.rochester.edu/smd/mbi/xenopus/Protocols/FV3ELISA.pdf

Category: ELISA Protocols

Protocol for ELISA for detection of Xenopus antibodies against FV3. - [Read ELISA for Detection of Xenopus Antibodies Against FV3 Protocol]

ELISA for Detection of Xenopus Antibodies Against FV3 Protocol - http://www.urmc.rochester.edu/smd/mbi/xenopus/Protocols/FV3ELISA.pdf

Category: Xenopus Protocols

Protocol for ELISA for detection of Xenopus antibodies against FV3. - [Read ELISA for Detection of Xenopus Antibodies Against FV3 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]

Flow Cytometry Analysis Protocol - http://www.cbrinstitute.org/labs/springer/protocols/qing_facs.html

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Sorting Protocols

Flow cytometry is a widely used method for characterizing and separating individual cells. This basic protocol focuses on: measure fluorescence intensity produced by fluorescent-labled antibodies and ligands that bind specific cell-associated molecules. Includes: Immunofluorescence Staining and Flow Cytometry Analysis. - [Read Flow Cytometry Analysis Protocol]

Flow Cytometry Analysis Protocol - http://www.cbrinstitute.org/labs/springer/protocols/qing_facs.html

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Flow cytometry is a widely used method for characterizing and separating individual cells. This basic protocol focuses on: measure fluorescence intensity produced by fluorescent-labled antibodies and ligands that bind specific cell-associated molecules. Includes: Immunofluorescence Staining; Flow Cytometry Analysis. - [Read Flow Cytometry Analysis Protocol]

Fluorescent Dyes for Flow Cytometric Analysis - http://www.ebioscience.com/ebioscience/appls/Dyes.htm

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

Fluorescent dyes absorb light at certain wavelengths and in turn emit their fluorescence energy at a higher wavelength. Each dye has a distinct emission spectrum, which can be exploited for multicolor analysis. eBioscience antibodies are available conjugated to a wide variety of fluorochromes. - [Read Fluorescent Dyes for Flow Cytometric Analysis]

Functional Analyses for Site-Specific Phosphorylation of a Target Protein in Cells - http://www.natureprotocols.com/2007/01/10/functional_analyses_for_sitesp.php

Category: Antibody Protocols: Antibody Production Protocols

Novel strategy of immunizing a phosphorylated peptide or a synthetic phosphopeptide, which corresponds to the protein phosphorylated at a targeted residue. Method has been applied to the production of antibodies that can specifically recognize the other types of site-specific protein modification, such as acetylation, methylation, and proteolysis. - [Read Functional Analyses for Site-Specific Phosphorylation of a Target Protein in Cells]

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.