Well, the answer is not a short one. Rabbit monoclonal antibodies (RabMAbs) have shown to play an important role as research and diagnostic reagents. One area where RabMAbs have demonstrated their superior performance is in immunohistochemical analysis of formalin fixed paraffin-embedded tissues based on their high affinity and sensitivity.
Epitomics has developed a large number of high quality rabbit monoclonal antibodies against difficult targets such as protein phosphorylation sites.
Rabbit monoclonal antibodies (RabMAb) combine the advantage of high affinity, attributable to their rabbit origin, and high specificity due their monoclonal nature.
The general procedure for making a rabbit monoclonal is very similar to the conventional mouse monoclonal. Making rabbit monclonals has proven much more difficult than mouse monoclonals which have been around since the late 1970's (1). In fact no one had successfully made a rabbit monoclonal antibody until the work by Dr. Katherine Knight in 1995 (2). Since the work in 1995 and subsequent work by others (3, 4, 5), and with the availability of the fusion partner, Epitomics obtained the exclusive rights to the technology enabling the creation of rabbit hybridomas. The following year, Dr Robert Pytela and his colleagues established a robust system for the development of rabbit monoclonal antibodies. Using this proprietary RabMAb technology, Epitomics has successfully produced a large number of antibodies which perform well in immunohistochemical analysis of formalin fixed paraffin-embedded tissues.
The Epitomics technology page in their web site offers an excellent general overview of the manufacturing procedure.
Rabbit antibodies are generally high affinity and are an ideal species to choose for making antibodies against mouse and rat proteins as many antibodies made in mice or rats will not react against mouse or rat proteins (2). From Epitomics’ experiences, the rabbit immune system appears to be capable of generating affinities against small epitopes which are not recognized by the mouse.
Rabbit polyclonal antibodies have successfully been used as research and even diagnostic applications for many years. However the inherent benefits of monoclonal antibody production quality and reliability have not been available for rabbit antibodies until now. Epitomics RabMab technology thus expands the antibody repertoire available for potential research, diagnostic and therapeutic applications.
RabMab affinities in the picomolar range, 10-12 Kd (M) are possible. Typical affinities are in the 10-9 10-10 Kd (M) range.
RabMabs can be used in the same applications as any antibody . RabMabs have been used successfully in western analysis, immunocytochemistry, imunohistochemistry, immunoprecipitation and flow cytometry. A c-kit rabbit monoclonal antibody was also recently approved for use in a diagnostic application. As with any antibody the use of a specific antibody should be validated and a given antibody may not be appropriate for use in every application. A list of recommended applications for each of Epitomics RabMab's can be found with the product specific information in the Epitomics web site.
Benefits of RabMAbs Technology
RabMAbs combine the inherent advantages of the Rabbit immune system with the specificity and consistency of a monoclonal antibody as rabbits provide a superior immune response
• More diverse range of epitopes recognized by Rabbits. Rabbit monoclonal technology enables the development of monoclonals against antigens that were previously difficult or unobtainable using the Mouse system.
• Rabbits respond well to immunogens which are not very immunogenic in mice -peptides, small molecules, phosphorylationsites etc.
• Epitomics RabMAbs are at the high end of typical antibody affinities. Randomly chosen RabMAbs from three separate projects show extremely high affinities (as measured by BIA core analysis).
• Ability to use mouse specific antigens. Rabbit mAbs can be raised against murine specific targets –especially useful for mouse animal model studies.
• RabMAbs have high affinity and specificity. Epitomics rabbit monoclonal technology has proven to be very robust with a 90% success rate with customer targets.
RabMAbs are suitable for use in multiple applications:
• Western blot analysis and Immunoprecipitation
• Immuno-fluorescence
• IHC (paraffin embedded tissues)
• Flow Cytometry
Hope this helps,
Sincerely,
Ronald
Refer a Friend
Link To Us
Bookmark Us