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ABH (p-Azidobenzoyl hydrazide) PDF - http://www.piercenet.com/files/0248dh4.pdf

Category: Molecular Biology Protocols: Carbohydrate Protocols

The ABH crosslinker binds to the Fc portion of an antibody molecule, away from the antigen binding site, resulting in a divalent immunologically active immunoglobulin derivative. Pierce - [Read ABH (p-Azidobenzoyl hydrazide) PDF]

Analysis of Proteins by Immunoprecipitation - http://dps.ufl.edu/hansen/protocols/imp98.prt.htm

Category: Protein Protocols: Immunoprecipitation Protocols

Antibody-antigen complexes are removed from solution by addition of an insoluble form of an antibody binding protein such as Protein A, Protein G or second antibody. Immunoprecipitation protocols / methodology and technical background information. P.J. Ha - [Read Analysis of Proteins by Immunoprecipitation]

Antibody Addition and Detection for Staining Caenorhabditis elegans Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4523

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

Extreme care should be used to identify and verify positive reactions, however, because cross-reactions are common. Counterstaining is essential for examining worms by immunofluorescence and is used to identify the exact cell in which an antigen appears. Methods for counterstaining include labeling all cells with a fluorescent dye that is specific for nucleic acids (e.g., DAPI or propidium iodide) and using GFP driven by tissue-specific promoters. - [Read Antibody Addition and Detection for Staining Caenorhabditis elegans Protocol]

Antibody Array Production and Analysis - http://www.stanford.edu/group/robinsonlab/protocols/protocols_tools.htm

Category: Antibody Microarray Protocols

Antibody Array Production and Data Analysis. Probing Antigen Arrays and washing/ Blocking. Great Protocols and information. Brian Kidd. Stanford. - [Read Antibody Array Production and Analysis]

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]

Antigen Design and Sera Purification - http://www.sigmaaldrich.com/Brands/Sigma_Genosys/Custom_Antisera/Key_Resources/Antigen_Design.html

Category: Immunology: Antigen

Antigen Design and Sera Purification. Custom antisera. Sigma Aldrich. Peptide Selection and Design, Coupling Strategy # Selecting the Protein Carrier, Multiple Antigenic Peptides (MAPs), Choice of Host, Adjuvant, Immunization, & Sera Collection, Antisera Purification, Ammonium Sulfate Precipitation, Protein A/G, Immunoaffinity Purification. - [Read Antigen Design and Sera Purification]

Antigen Preparation for Polyclonal Antibody Production in Rabbits - http://researchadmin.asu.edu/compliance/iacuc/sop/RabbitantibodyproductionSOP.doc

Category: Immunology: Antigen

Antigen Preparation for Polyclonal Antibody Production in Rabbits - [Read Antigen Preparation for Polyclonal Antibody Production in Rabbits]

Antigen Retrieval - Immunohistochemistry - http://axon.med.harvard.edu/~cepko/protocol/mike/A2.html

Category: Immunology: Antigen Retrieval

Denaturing Solution and Procedure for AR. Mike Dyer. - [Read Antigen Retrieval - Immunohistochemistry]

Antigen Retrieval for immunohistochemistry - http://herpesvirus.tripod.com/research/protoc.htm#Antigen%20Retrieval%20for

Category: Immunology: Antigen Retrieval

Antigen Retrieval for immunohistochemistry. Microwave method, Autoclave method. slides and staining. Janos Luka. Eastern Virginia. - [Read Antigen Retrieval for immunohistochemistry]

Antigen Retrieval for Immunohistochemistry with Paraffin-Embedded Tissues Protocol - http://www.animal.ufl.edu/hansen/Protocols/Immunohistochemistry_paraffin.htm

Category: Immunohistochemistry Protocols: Antigen Retrieval Protocols

Protocol used to for immunohistochemistry on paraffin-embedded sections. Based on use of microwave energy to effect antigen retrieval. The immunohistochemistry procedure, is for use of Biomeda's HistoScan kit based on a streptavidin-peroxidase/biotinylated second antibody detection system with 3-amino, 9-ethylcarbazole (AEC) as chromogen. Undoubtedly, other kits or home-made reagents will also work . - [Read Antigen Retrieval for Immunohistochemistry with Paraffin-Embedded Tissues Protocol]

Antigen Retrieval PDF - http://www.dakousa.com/ishbantigenretr.pdf

Category: Immunology: Antigen Retrieval

Antigen Retrieval. Principles and Technique. A handbook of Immunochemical Staining Methods and techniques. Marc Key - [Read Antigen Retrieval PDF]

Antigen Retrieval Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/A2.html

Category: Immunohistochemistry Protocols: Antigen Retrieval Protocols

Protocol for antigen retrieval. - [Read Antigen Retrieval Protocol]

AR Protease Digestion Method - http://www.chemicon.com/techsupp/Protocol/Protease.asp

Category: Immunology: Antigen Retrieval

Protease Antigen Retrieval (Trypsin) Protocol. Chemicon. - [Read AR Protease Digestion Method]

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]

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]

Combined DNA In Situ Hybridization and Immunocytochemistry Protocol - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_127-134.pdf

Category: Cytogenetics Protocols: In Situ Hybridization Protocols

Protocol for combined DNA in situ hybridization and immunocytochemistry for the simultaneous detection of nucleic acid sequences, proteins, and incorporated BrdU in cell preparations. Includes: Cell preparations and BrdU labeling; Detection of antigen by immunocytochemistry (ICC); Visualization of ICC antigen; -Gal-BCIG reaction (for producing a blue precipitate visible under brightfield microscopy); Cell processing for in situ hybridization; In situ hybridization (ISH); etc... - [Read Combined DNA In Situ Hybridization and Immunocytochemistry Protocol]

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

Category: Antibody Protocols: Antibody Labeling Protocols

Protocol for the detection of autoantibodies with self-assembling radiolabeled antigen tetramers. - [Read Detection of Autoantibodies with Self-Assembling Radiolabeled Antigen Tetramers 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]

Double immunofluorescence staining for BCL-6 - http://icg.cpmc.columbia.edu/cattoretti/Protocol/Immunohistochemistry/DoubleIFstain.html

Category: Molecular Imaging: Immunofluorescence Microscopy

Double immunofluorescence staining for BCL-6. unfixed or acetone-fixed specimens. dewaxed, antigen retrieved slides - [Read Double immunofluorescence staining for BCL-6]

Double Immunofluorescence Staining for BCL-6 Protocol - http://icg.cpmc.columbia.edu/cattoretti/Protocol/immunohistochemistry/DoubleIFstain.html

Category: Immunology: Immunofluorescence Protocols: Immunofluorescence Staining Protocols

Protocol for double immunofluorescence staining for BCL-6. Any type of tissue is suitable for this technique, as long as the antigenicity for your antigen(s) is preserved. Includes: unfixed or acetone-fixed specimens; dewaxed, antigen retrieved slides. - [Read Double Immunofluorescence Staining for BCL-6 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]

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]

Articles (1-3 of 3)

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.

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