A METHOD FOR THE DETERMINATION OF INORGANIC-PHOSPHATE IN THE PRESENCE OF LABILE ORGANIC PHOSPHATE AND HIGH-CONCENTRATIONS OF PROTEIN - APPLICATION TO LENS ATPASES
A METHOD FOR THE DETERMINATION OF INORGANIC-PHOSPHATE IN THE PRESENCE OF LABILE ORGANIC PHOSPHATE AND HIGH-CONCENTRATIONS OF PROTEIN - APPLICATION TO LENS ATPASES
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DOI：
10.1016/0003-2697(88)90002-4
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发表时间：
1988-01-01
影响因子：
2.9
通讯作者：
TOLOSA, S
中科院分区：
文献类型：
--
作者：
CHIFFLET, S;TORRIGLIA, A;TOLOSA, S
本文对传统的磷酸盐测定方法进行了改进。该方法具有高灵敏度，1 nmol Pi的吸光度变化为0.060 A850单位。该方法重现性好，颜色至少稳定3.5 h。此外，十二烷基硫酸钠的使用使得可以在没有有机磷酸盐水解的情况下停止酶促反应。它还显示了不同来源的高浓度溶液的干扰极低。特别令人感兴趣的是其对蛋白质的高耐受性，允许样品中多达50 mg/ml的人血清蛋白而没有沉淀或颜色干扰。由于这些原因，它被证明在测定组织如具有低比活性的眼透镜中的ATP酶中非常有用。
We present here an improvement in the classical molybdate method for inorganic phosphate determination. This method has high sensitivity, with 1 nmol of Pi giving an absorbance change of 0.060 A850 unit. It is highly reproducible and the color remains stable for at least 3.5 h. In addition the use of sodium dodecyl sulfate makes it possible to stop enzymatic reactions without organic phosphate hydrolysis. It also shows an extremely low interference by highly concentrated solutions of different origins. Of special interest is its high tolerance to protein, permitting as much as 50 mg/ml of human serum protein in the sample without precipitation or color interference. For these reasons, it proves to be very useful in the determination of ATPases in tissues such as the ocular lens with low specific activity.