Related Experiment Video

Updated: Sep 28, 2026

Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography
10:50

Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography

Published on: March 10, 2010

Isolation of the I-Ak core complex and its associated gamma-polypeptide by affinity chromatography on monoclonal

R Zecher, K Reske

    Molecular Immunology
    |August 1, 1982
    PubMed

    Spleen cell-derived I-Ak antigens have previously been shown by us to be comprised of four noncovalently associated polypeptide chains (alpha, gamma, beta 1, beta 2). Here we report that pretreatment of the detergent solubilized four-chain structure with chaotropic ions yielded an alloantigenically reactive core complex (alpha/beta 1, beta 2) and a dissociated polypeptide gamma-chain which was not recognized by monoclonal antibody 10-2.16 (anti-I-Ak). The lack of alloantigenic determinants on gamma and the observation that gamma did not reassociate with alpha/beta 1, beta 2 upon removal of the chaotropic reagent suggested a rapid purification procedure for both moieties. A pure preparation of the serologically intact core complex was achieved by repeated affinity chromatography on hybridoma immunoadsorbent 10-2.16 Sepharose CL-4B. Concurrently its pertinent gamma-polypeptide was obtained in a highly enriched form and was purified to homogeneity by an additional gel electrophoretic step.

    More Related Videos

    Isolation and Characterization Of Chimeric Human Fc-expressing Proteins Using Protein A Membrane Adsorbers And A Streamlined Workflow10:33

    Isolation and Characterization Of Chimeric Human Fc-expressing Proteins Using Protein A Membrane Adsorbers And A Streamlined Workflow

    Published on: January 8, 2014

    Purification of Native Complexes for Structural Study Using a Tandem Affinity Tag Method10:36

    Purification of Native Complexes for Structural Study Using a Tandem Affinity Tag Method

    Published on: July 27, 2016

    Related Concept Videos

    Overview Of Cell Separation And Isolation01:20

    Overview Of Cell Separation And Isolation

    Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.
    Immunoprecipitation01:20

    Immunoprecipitation

    Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
    Chromatin Immunoprecipitation
    Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
    Affinity Chromatography01:03

    Affinity Chromatography

    Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...