
%0 Journal Article
%A Jones, WT 
%A Jones, SD 
%A Liddane, CP 
%A Reynolds, PHS 
%T Measurement of Aspartate Aminotransferase-P<sub>2</sub> in Developing Lupin Nodules Using a Two Site ELISA Employing Monoclonal Antibodies
%J Functional Plant Biology
%D 1992/04/01
%V 19
%N 2
%P 147-153
%K 
%R https://doi.org/10.1071/PP9920147
%U https://www.publish.csiro.au/paper/PP9920147 
%X Leguminous root nodules contain two forms of aspartate aminotransferase (AAT). AAT-P1 is con- stitutively expressed in roots and nodules, whereas the second isoenzyme, AAT-P2, is induced in developing nodules. A two site ELISA system was developed to quantify AAT-P2 in nodule extracts. The assay comprised two affinity-purified monoclonal antibodies directed against different epitopes on the AAT-P2 molecule. The sensitivity was less than 200 pg mL-1, and AAT-P2 could be quantified in the range 1 to 15 ng mL-1. The assay was specific for AAT-P2, and did not measure AAT-P1. The assay was used to specifically measure AAT-P2 in the developing legume root nodule formed by the symbiotic liaison between Rhizobium lupini NZP2257 and the roots of Lupinus angustifolius L. The ELISA method, which measures AAT-P2 protein, was contrasted with an approach which relied on the heat lability of AAT-P2 at 60&#176;C to measure individual isoenzyme catalytic activity. 
