Salto R, Oliver J, del Mar Sola M, Vargas A M
Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de Granada, Spain.
Kidney Int. 1991 Jun;39(6):1162-7. doi: 10.1038/ki.1991.147.
Antibodies against purified rat-kidney-cortex pyruvate carboxylase were raised in rabbits. These polyspecific antibodies recognize pyruvate carboxylase enzymes alone without cross reactivity with other carboxylases as detected by immunoblotting. The abundance of pyruvate carboxylase in the various renal fractions was measured by ELISA and its activity by the fixation of [14C]CO3H-. The results were corroborated by a combination of immunocytochemistry and transmission electron microscopy. A good correlation was found between the enzymatic activity and the quantity of enzyme contained in each fraction. The kidney-cortex pyruvate carboxylase was primarily located in proximal tubules, in accord with its important role in gluconeogenesis.
用纯化的大鼠肾皮质丙酮酸羧化酶免疫家兔制备抗体。这些多特异性抗体仅识别丙酮酸羧化酶，通过免疫印迹检测，它们与其他羧化酶无交叉反应。通过酶联免疫吸附测定法（ELISA）测量各肾部分中丙酮酸羧化酶的丰度，并通过固定[14C]CO3H-来测定其活性。免疫细胞化学和透射电子显微镜相结合的方法证实了这些结果。发现酶活性与各部分所含酶的量之间具有良好的相关性。肾皮质丙酮酸羧化酶主要位于近端小管，这与其在糖异生中的重要作用一致。
