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Interleukin-12 is a cytokine that plays a central role in mediating cell mediated immunity via enhancement of a TH1 cell response. IL-12, unusually for a cytokine, has a heterodimeric structure made up of 35 kDa and 40 kDa subunits. The aim of this study was to produce and characterize monoclonal antibodies to recombinant human IL-12. Twenty-two monoclonal antibodies to IL-12 were successfully produced and subunit specificity determined using recombinant human IL-12 and chimeric murine/human IL-12. All antibodies were shown to react with the p40 subunit by ELISA and immunoblotting with three of the MAbs being found to cross-react with murine IL-12. Using two individual bioassays for IL-12, seven of the MAbs were shown to neutralize biological activity of IL-12. Ten of the antibodies were found to be of use in immunocytochemistry, reacting with LPS-stimulated peripheral blood monocytes. The approaches and difficulties encountered in characterizing antibodies to a heterodimeric cytokine are discussed together with possible reasons for the failure to generate antibodies to the p35 subunit.
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P M Patel, and
A J Stanley, and
E Ingham, and
M Wadhwa, and
C Bird, and
R Thorpe, and
P J Selby, and
R E Banks
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R W Carter, and
P M Patel, and
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E Ingham, and
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C Bird, and
R Thorpe, and
P J Selby, and
R E Banks
January 2015,
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R W Carter, and
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R Thorpe, and
P J Selby, and
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February 1988,
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R W Carter, and
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R E Banks
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R W Carter, and
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R E Banks
October 2009,
Hybridoma (2005),
R W Carter, and
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E Ingham, and
M Wadhwa, and
C Bird, and
R Thorpe, and
P J Selby, and
R E Banks
March 2011,
Veterinary immunology and immunopathology,
R W Carter, and
P M Patel, and
A J Stanley, and
E Ingham, and
M Wadhwa, and
C Bird, and
R Thorpe, and
P J Selby, and
R E Banks
December 2006,
Tissue antigens,
R W Carter, and
P M Patel, and
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E Ingham, and
M Wadhwa, and
C Bird, and
R Thorpe, and
P J Selby, and
R E Banks
May 2005,
The Journal of veterinary medical science,
R W Carter, and
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C Bird, and
R Thorpe, and
P J Selby, and
R E Banks
November 1991,
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R W Carter, and
P M Patel, and
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E Ingham, and
M Wadhwa, and
C Bird, and
R Thorpe, and
P J Selby, and
R E Banks
August 1987,
Hybridoma,
