|Product Includes||Quantity (with Count)||Storage Temp|
|Malachite Green Reagent||2 x 28 ml||RT|
|Phosphate Standard (1 mM)||1 x 0.5 ml||RT|
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
|Volume (µl) of diluted stock (0.1mM)||160||140||120||100||80||60||40||20||10||5||0|
|Volume (µl) of distilled water or diluent||90||110||130||150||170||190||210||230||240||245||250|
|Phosphate [pmol] per 25 µl||1600||1400||1200||1000||800||600||400||200||100||50||0|
NOTE: Malachite Green is a highly sensitive phosphate detection solution. Make sure any reagent used in the assay is free of inorganic phosphates, as this will greatly increase the background absorbance of the assay. Enzyme reactions should be performed in a final volume of 25 µl (the same volume as that used for the standard curve). Before proceeding, confirm that your enzyme samples are free of contaminating inorganic phosphate. To determine phosphate contamination, add Malachite Green Reagent (100 µl) to 2-5 µl of sample. Any phosphate contamination will result in a color change to green. Measure absorbance at 620-660 nm and compare to standard curve to determine amount of contaminating inorganic phosphate. If required, contaminating inorganic phosphate can be removed using a desalting column or dialysis.
posted June 2020
Protocol Id: 2058
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