Phosphate detection

Catecholamines, nerve transmitters monitored in brain and heart patients, are separated on C18 using octane sulfonate ion pairing in 6% An/water (pH 3) with added EDTA and phosphate. Detection can be at UV, 270 nm, or by electrochemical detection at +0.72 V for maximum sensitivity. Other tyrosine and tryptophan metabolite neurotransmitters such as serotonin, VMA, and HMA can be analyzed with ion pairing and EC detection. 

The use of (dppe)Pt S2C2(2-py)(CH2CH2OH) in phosphate detection is further complicated by the nature of the emissive states. The triplet emission of... 

Subject the other dried chromatogram to one of the following phosphate-detection proce-... 

Fig. 8 Malachite green assay for phosphate detection. The enzymatic reaction produces inorganic phosphate P. A secondary reaction is used to form a phosphomolybdate-malachite green complex, which has a strong absorbance at 660 nm.

The classical phosphate detection method originally developed by Bell and Doisy [3] and further optimized by Fiske and SubbaRow... 

Some properties of phosphopeptides make them preferable to the native phosphoprotein substrates for use with phosphate detection systems. The values of for peptide substrates are two to three orders of magnitude larger than for protein substrates and allow setting assays with an appropriate substrate concentration using standard phosphate detection approaches. In addition, short synthetic peptides are inexpensive and easy to obtain. Nonetheless, although phosphopeptide substrates are clearly useful in exploring interactions in the immediate vicinity of the phosphatase active site, they are unable to probe distant (allosteric) sites. 

Key words ENPPl, ATP, ADP, Pyrophosphatase, Phosphodiesterase, Phosphate detection,... 

PiColorLock Phosphate Detection System (Innova Biosciences, Cambridge, UK) 100 pM phosphate standard. Gold reagent solution, accelerant solution, and stabilizer solution. Store at 4 °C or on ice during assay. 

HTS Protocol for Using pNPP and Free Phosphate Detection with the Biomol Green Reagent... 

Fig. 11.1.4. Separation of uracil and 5-fluorouracil bases, nucleosides and nucleotides by reversed phase ion-pair HPLC. Chromatographic conditions column, Bondapak Cig (300 x 4 mm) mobile phase, (from 0-30 min) 0.1 mM tetrabutylammonium hydrogen sulphate (Cjg), 2.5 mM tetraethylammonium bromide (Cg) and 2% methanol in 2 mM sodium acetate, 1.5 mM phosphate buffer, pH 6.0 (Buffer A) (from 30-50 min) Buffer A-i-30 mM phosphate detection, UV at 254 nm. Peaks FU, fluorouracil FUR, fluorouracU riboside/ FUdR, fluorouracil deoxyriboside FUMP, fluorouridine 5 -monophosphate 5 dFUR, 5 -deoxyfluorouracil riboside FdUMP, deoxyfluorouri-dine monophosphate UDPG, uridine diphosphoglucose UDP, uridine diphosphate dUDP, deoxyuridine monophosphate UTP, uridine triphosphate. Reproduced from Au et al. (1982), with permission.

Carbon monoxide phosphatase determination lndol-3-yl sodium phosphate phosphate detection Ammonium molybdate (VI)... 

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