Detection assay for potassium ions using a potassium-dependent urea amidolyase enzyme
The invention relates generally to the field of potassium ion detection. In particular, the invention provides kits for assaying for potassium ions in a sample using a potassium-dependent urea amidolyase (UAL) enzyme.
1. A kit for assaying for potassium ion concentration in a biological sample, which kit comprises
a) a first composition comprising a potassium-dependent urea amidolyase, wherein the amidolyase consumes urea and forms inorganic phosphate (P i ); and
b) means for assaying the urea consumed or the P i formed by the urea amidolyase to determine the presence or amount of the potassium ions in the sample, whereby the potassium ion concentration is determined.
2. The kit of claim 1 , wherein the first composition further comprises glycogen, phosphorylase a, oxidized β-nictinamide adenine dinucleotide (NAD), phosphoglucomutase, glucose-6-phosphate dehydrogenase (G-6-PDH), 2-(4-iodophenyl)-3-(4-nitrophenyl)-5(2,4-disulfophenyl)-2H-tetrazolium (WST-1), and 1-methoxy-5-methyl-phenazin methyl sulfate (PMS), wherein the means for assaying the urea consumed or the P 1 formed if potassium ions are present is a reduction of WST-1 in the presence PMS to form formazan.
3. The kit of claim 2 , further comprising a second composition comprising urea, adenosine triphosphate (ATP), and MgCl 2 .
4. The kit of claim 3 , wherein the second composition further comprises a protein.
5. The kit of claim 4 , wherein the protein is bovine serum albumin.
6. The kit of claim 3 , wherein the second composition further comprises a buffer.
7. The kit of claim 6 , wherein the buffer comprises NaHCO 3 .
8. The kit of claim 1 , wherein the kit further comprises standards for calibration of the assay.
9. The kit of claim 8 , wherein the standards for calibration are a low potassium serum standard and a high potassium serum standard.