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An immunological method for quantitative determination of photosynthetic fructose-1,6-bisphosphatase in leaf crude
R Hermoso1, A Chueca, J J Lazaro
1Unidad de Bioquímica Vegetal, Estación Experimental del Zaidín (CSIC), Profesor Albareda 1, 18008, Granada, Spain.
Abstract:
An immunological method for quantitative determination of photosynthetic fructose-1,6-bisphosphatase in crude extracts of leaves is proposed. It is based on the ELISA technique, and offers two modifications. A non-competitive technique has a higher sensitivity and is the right option for samples of low fructose-1,6-bisphosphatase content. However, this method is not sufficiently specific when the total protein is higher than 5 μg/cm(3); so, despite its lower sensitivity, in these circumstances a competitive technique is more suitable. Thus photosynthetic fructose-1,6-bisphosphatase can be measured without interferences from the gluconeogenic cytosolic enzyme of the photosynthetic cell or from a non-specific phosphatase present in the chloroplast.
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