A method for the determination of inorganic phosphate in the presence of labile organic phosphate and high concentrations of protein: Application to lens ATPases

✍ Scribed by S. Chifflet; A. Torriglia; R. Chiesa; S. Tolosa

• Publisher: Elsevier Science
• Year: 1988
• Tongue: English
• Weight: 269 KB
• Volume: 168
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(88)90002-4

✦ Synopsis

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 &a 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 m&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. o 1988 Academic ~rw., Inc.