Protocol for: Antibody Purification
Link to Full Protocol:
Abstract or Description:
Hybridoma technology is a technology of forming hybrid cell lines (hybridomas) by fusing a specific antibody-producing B cell with a myeloma (B cell cancer) cell that is selected for its ability to grow in tissue culture and for an absence of antibody chain synthesis. The antibodies produced by the hybridoma are all of a single specificity and are therefore monoclonal antibodies.
Creative BioLabs provides a full range of services in custom polyclonal and monoclonal antibody production from gene expression or peptide synthesis to antibody purification and labeling. We have the capacity of producing antibodies in either research quantities or in large scale for diagnostic industry. We do epitope prediction, peptide synthesize, protein expression, polyclonal or monoclonal antibody production as well as antibody purification from cell culture, ascites fluid or whole serum. We also prepare mono-specific polyclonal antibodies by ligand affinity chromatography. Importantly, we can produce monoclonal antibodies within 70 days. Creative BioLabs also has the expertise in handling antigen insufficiency and targeting very impure antigens.
Creative BioLabs has established standard operating procedures in selectively identifying antibodies that bind not only to immunogens but also to the corresponding target antigens in ELISA, Western blotting, Immuno-precipitation, Immuno-fluorescence and Immunohistochemistry. Frequently, polyclonal or monoclonal antibodies raised using peptides and recombinant proteins cannot recognize the target antigens, e.g. a protein antigen in tissue, and are essentially useless. To generate antibodies that recognize a native antigen, we employ or predict the 3-D structure of the target protein and design peptide immunogens that display on the protein surface and contain linear continuous epitopes. If a recombinant protein immunogen is used, we usually express it in a soluble form to retrieve the immunogenic properties of its native counterpart. Finally, we use our well-tested ELISA, Western blotting, IP, IF and IHC screening protocols to validate (monoclonal) antibodies that recognize the natural target antigen.
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Posted By: candyswift on 10/13/2015 7:23:15 AM