Hello and thanks

In this case, I do not have any information.

sincerely

Malachite Green/molybdate reagents for inorganic phosphate detection can detect phosphate concentrations as low as about 1 µM, although the sensitivity varies depending on the formulation. Such high sensitivity makes it imperative that sources of phosphate contamination be eliminated from the assay. Only the purest water (Milli-Q water, preferably) should be used to prepare solutions. Be careful about glassware because soap and tap water residues can contain phosphate. It is preferable to use only new plasticware.

If you find Malachite Green reagents to be too sensitive for your application, consider switching to a less sensitive molybdate-only phosphate detection reagent.

yes, I agree with , this kits is too sensitive, I am using mili-Q water and it is very good with no color developed.  Didi you used this kits for Adenosine synthase enzyme or Nucleotidase?

Hi, I haven't used any kit, but home-made solutions and it worked fine. At that time we didn't even have MilliQ water, so that shouldn't be so crucial.

However, as Adam wrote, you must use either plasticware or rinse the glassware with concentrated HCl. That worked for me.

Further, you must strip your buffers and other reagents of anything that is not necessary.  Although glycerol, as I'm just now looking, actually helped.

Awesome, so how I can prepare home made solution, and what the importance of glycerol? is it useful and if so at what conc. you have used ?

From my bachelor thesis:

73.12 ml of solution A (0.045% malachite green in water) was mixed with 24.38 ml

of solution B (4.5% ammonium heptamolybdate in 4 M HCl) while vortexing to avoid

precipitation. After 20 minutes, 2.5 ml of solution C (1.5% Tween in water) was added.

This working reagent is stable for at least 24 hours.

One reaction volume was added to the IPT reaction (as described before), mixed

and incubated for 40 minutes at room temperature. Afterward, absorbance at 620 and

636 nm was read against blank (see below). Blank was set up equally as reaction, only DMAPP was omitted.

The glycerol (fin. conc. 20%) was important for my enzyme, because it was not much stable.

thanks Tomas,you was working on this enzyme adenosine synthase or Nucleotidase?