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High Throughput Colorimetric ATPase Assays (inc. PiColorLock)

Key Features of PiColorLock™ - phosphate detection reagent:

 

  • Colorimetric Assay – competitor assays are radioactive.
  • Special additive ensures low backgrounds with acid-labile substrates.
  • Unparalleled stability of phosphate-dye complexes.
  • Reagent is compatible with almost any assay buffer.
  • No inhibition of color development by high concentrations of protein.
  • Stable reagent formulation - long shelf life.

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Introduction

Colorimetric assays for ATPases are invariably based on the formation of colored complexes between an inorganic phosphate and a dye molecule under acidic conditions.

picolorlock

Competitor assays are beset with several problems including reagent precipitation.  At the heart of Innova Biosciences assay kit is PiColorLock™ Gold a non-radioactive, superior phosphate detection reagent which ensures high stability of the colored dye-phosphate complexes (green color- see below).

cuvettesaltered web

PicolorLock™ Gold has also been designed to:

  • Prevent background signals arising from non-enzymatic (i.e. acid) hydrolysis of the substrate. The figure below shows ATP incubated in three detection reagents. A steadily rising background signal is seen with competitor reagents, whereas PiColorLock™ gives baseline readings.

assay background web

  • Have a large linear range, thus reducing the need for sample dilution.

linear range web

Together with specially purified substrates and proprietary stabilizers Innova Biosciences kits offer the lowest possible assay background and outstanding assay performance.

PiColorLock™ Gold can also be purchased for developing simple assays for any phosphate-generating enzyme.

For more information, please watch our webinar on drug screening-assays for phosphate-generating enzymes below:

 

Related Reagents

PiBind™ resin provides a quick and easy way to remove contaminating Pi from buffers and from samples derived from cells and tissues.

by InnovaBiosciences 

I have a high background in my ATPase assay and I definitely do not have free phosphate in my sample

This is almost always due to inadequate mixing of the special stabiliser with the sample and detection reagent. Make sure the stabiliser is pipetted up and down several times to ensure thorough mixing.

I would like to measure the conversion of pyrophosphate to phosphate. Can I use the PiColorLock Gold Phosphate Detection System for this purpose?

Yes, only the phosphate will give a signal; pyrophosphate will not.

I have 5% DMSO in my assay. Can I use PiColorLock Gold?

Yes, the reagent is designed for drug screening work and other situations that require DMSO.

I have phosphate in my enzyme. What can I do?

You can dialyse or desalt the enzyme into a phosphate-free buffer. Alternatively, you can use a special resin (PiBind) to remove the phosphate.

 

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192 assays (601-0120)

In stock

£234.00

480 assays (601-0121)

In stock

£458.00

5 x 480 assays (601-0122)

In stock

£1,915.00
Our shipping times are as follows:
48 hours to the US. 24 to 48 hours to Europe.

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