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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]

Making Antibodies to Synthetic Peptides Protocol - http://medicine.ucsd.edu/hypertension/list_of_protocols/16_antibodies_to_peptides.htm

Category: Antibody Protocols: Antibody Production Protocols

Protocol for making antibodies to synthetic peptides. - [Read Making Antibodies to Synthetic Peptides Protocol]

Making Monoclonal Antibodies Protocol - http://dicty.cmb.northwestern.edu/Chis_lab/Lab%20Manual/immunological_techniques.htm

Category: Antibody Protocols: Antibody Production Protocols

Protocol for making monoclonal antibodies. - [Read Making Monoclonal Antibodies Protocol]

Mitogen Activated Protein Kinase Guide - http://www.promega.com/guides/sigtrans_guide/strg_chap01.pdf

Category: Signalling Signal Transduction Protocols: MAPK Protocols

Mitogen Activated Protein Kinase Guide. MEK, MEKK, MAPK, MAPKK, antibodies and kits Promega. - [Read Mitogen Activated Protein Kinase Guide]

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]

Paraformaldehyde Fixation of Cells - http://www.biotech.iastate.edu/facilities/CELLHYB/Fixation.htm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Paraformaldehyde Fixation of Cells protocol. This fixation method is good for cells labeled by fluorochrome-conjugated antibodies to membrane antigens. It will stabilize the light scatter and labeling for up to a week in most instances, allowing you to be more flexible in scheduling cytometer time. Furthermore, it inactivates most biohazardous agents, so it is important from a safety standpoint as well. Iowa University. - [Read Paraformaldehyde Fixation of Cells]

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]

Preparation of Cells and Reagents for Flow Cytometry Protocols - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E661C3CC685613447CEC768412FDE0F&objectid=6674EA50F5C3D9A0189B376C7A89BF6B

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

A procedure for direct and indirect staining of single-cell suspensions of lymphoid tissue or peripheral blood lymphocytes to detect cell surface membrane antigens is presented. In addition, support protocols present methods for fluorescence labeling of purified antibodies. A protocol for flow cytometric analysis of intracellular antigens in single-cell suspensions is also included. - [Read Preparation of Cells and Reagents for Flow Cytometry Protocols]

Preparation Of Peptide-KLH Conjugates For Immunization - http://mitchison.med.harvard.edu/protocols/ab1.html

Category: Peptide Protocols: Antibodies from Peptides Protocols

Preparation Of Peptide-KLH Conjugates For Immunization. Coupling of Peptide to KLH. And generation of Antibodies using peptide-KLH conjugate - Claire Walczak. - [Read Preparation Of Peptide-KLH Conjugates For Immunization]

Preparation of Polyclonal Antibodies from Plasma or Serum Using the Gradiflow BF400 - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4236

Category: Antibody Protocols: Antibody Production Protocols

Polyclonal antibodies can be isolated from animal plasma or serum using the procedure described in this protocol. The Gradiflow BF400 instrument has two liquid streams that circulate through a separation cartridge positioned between two electrodes and composed of three hydrogel polyacrylamide membranes, which define the channels for the two sample streams. The central membrane forms a physical barrier between the two streams. - [Read Preparation of Polyclonal Antibodies from Plasma or Serum Using the Gradiflow BF400]

Probing Protein Interactions Using GFP and FRET Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3893

Category: Reporter Gene Assays: GFP Assay Protocols

In preparation for FLIM-FRET analysis, the appropriate donor and acceptor components must be introduced into live or fixed cells. The method of introduction depends on the nature of the components and the state of the cells. For example, plasmid DNAs encoding a protein of interest fused to a variant of GFP may be introduced into live cells by transfection or microinjection, whereas labeled antibodies are delivered by microinjection. - [Read Probing Protein Interactions Using GFP and FRET Protocol]

Probing Protein Interactions Using GFP and FRET. - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3960

Category: Reporter Gene Assays: GFP Assay Protocols

Describes how components (usually Fab fragments of antibodies) that are to be introduced into cells are labeled with fluorescent sulfoindocyanine dyes. - [Read Probing Protein Interactions Using GFP and FRET.]

Protein Coimmunoprecipitation in Arabidopsis Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4683

Category: Plant Biology Protocols: Arabidopsis Protein Peptide Protocols

Coimmunoprecipitation is one of the most useful techniques for revealing protein-protein interactions. Good negative controls to verify the specificity of the coimmunoprecipitation procedure are (1) performing the same immunoprecipitation experiment using beads coupled to the preimmune serum and (2) probing the Western blot with antibodies against protein known not to interact with the coimmunoprecipitated proteins under physiological conditions. - [Read Protein Coimmunoprecipitation in Arabidopsis Protocol]

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]

Purification of Lin-, c-kit+, Sca-1+ Bone Marrow Cells for Culture, Flow Cytometry, or Transplantaio - http://www.fhcrc.org/science/labs/fero/Protocols/Hematopoietic.pdf

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol for purification of Lin-, c-kit+, Sca-1+ bone marrow cells for Culture, Flow Cytometry, or Transplantaion. Includes: Antibodies for Lineage Depletion; Harvest Marrow; Lineage Depletion; Cell Sorting; Viral Transduction; Transplant; CFU assays; GFP and cell surface marker immunostaining. - [Read Purification of Lin-, c-kit+, Sca-1+ Bone Marrow Cells for Culture, Flow Cytometry, or Transplantaio]

Removal of Cross-reactive Antibodies from Antiserum: Affinity Chromatography Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3340

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

Protocol describes a method for removing antibodies that react with bacterially encoded proteins by passing a crude preparation of immunoglobulins through a column containing immobilized bacterial proteins. - [Read Removal of Cross-reactive Antibodies from Antiserum: Affinity Chromatography Protocol]

Removal of Cross-reactive Antibodies from Antiserum: Incubation with E. coli Lysate - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3767

Category: Antibody Protocols: Antibody Production Protocols

This protocol describes how antibodies that react with bacterial-encoded proteins may be removed from polyclonal antisera by incubation with a bacterial lysate. - [Read Removal of Cross-reactive Antibodies from Antiserum: Incubation with E. coli Lysate]

Removal of Cross-reactive Antibodies from Antiserum: Pseudoscreening Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3772

Category: Antibody Protocols: Antibody Production Protocols

Antibodies directed against bacterial and phage-encoded proteins can be removed from sera by incubation with nitrocellulose filters impregnated with lysates of uninfected or phage-infected E. coli. - [Read Removal of Cross-reactive Antibodies from Antiserum: Pseudoscreening Protocol]

Screening Expression Libraries Constructed in Plasmid Vectors Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4079

Category: DNA Protocols: cDNA Library Protocols

A cDNA library constructed in a plasmid expression vector of the pUC, pUR, or pEX series is plated on agar medium and then replicated onto filters, which are transferred to plates containing IPTG. After 2-4 hours of induction, the colonies are lysed with chloroform and then screened with appropriate antibodies. - [Read Screening Expression Libraries Constructed in Plasmid Vectors Protocol]

Storage of Ascitic Fluid Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4281

Category: Antibody Protocols: Antibody Handling Protocols

Ascitic fluid from B-cell-derived tumors is an extremely concentrated solution of specific antibodies. Ascitic fluid is often stored at -70ºC to prevent proteolytic degradation. - [Read Storage of Ascitic Fluid Protocol]

Storage of Purified Antibodies - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4282

Category: Antibody Protocols: Antibody Handling Protocols

This protocol describes the storage of "homemade" purified antibodies. Antibodies from commercial sources are normally supplied with proper storage conditions. - [Read Storage of Purified Antibodies]

Storage of Sera Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4279

Category: Antibody Protocols: Antibody Handling Protocols

This protocol describes the methods of storage for antibody-containing sera. Antibodies are resistant to a broad range of mildly denaturing conditions, so long-term storage is relatively easy. - [Read Storage of Sera Protocol]

T Cell Depletion by Cytotoxic Elimination Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E66365BA79D5D5A9DA4948DDF019E3B&objectid=6674981906D4B89C730A2E1ED1987295

Category: Immunology: Mononuclear Cell Isolation Protocols: T-Cell Isolation Protocols

Protocol describes removal of T cell subsets by cytotoxic elimination using CD-specific antibodies. Describes the complete removal of T cells from lymphocyte preparations based on the presence of the glycoprotein Thy-1 on the cell surface of T lymphocytes. Cytotoxic elimination is employed; however, Thy-1-specific antibodies are used rather than MHC class II-specific antibodies so that T cells are eliminated rather than B cells and accessory cells. - [Read T Cell Depletion by Cytotoxic Elimination Protocol]

T Cell Enrichment by Cytotoxic Elimination of B Cells and Accessory Cells Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E66364CDDD21842657A5068627A2F0C&objectid=667495E7DA47001CA835F427F96D1C3B

Category: Immunology: Mononuclear Cell Isolation Protocols: T-Cell Isolation Protocols

Describes T cell enrichment using cytotoxic antibodies, and also describes the depletion of T cells and their subpopulations using the same approach. In the latter unit, T cell surface markers (Thy-1, CD4, and CD8) are targeted by the cytotoxic antibodies. - [Read T Cell Enrichment by Cytotoxic Elimination of B Cells and Accessory Cells Protocol]

Tips and hints for the storage of antibodies - http://www.sysy.com/faqs/storage.html

Category: Protein Protocols: Western Blot Protocol: Antibody Storage Conditions

Tips and hints for the storage of antibodie. Storage of lyophilized antibodies, - [Read Tips and hints for the storage of antibodies]

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.