PiColorLock™ Phosphate Detection System
PiColorLock™ phosphate detection reagent
PiColorLock™ phosphate detection reagent changes colour in the presence of inorganic phosphate (Pi). This can be exploited to measure any enzyme that generates Pi including ATPases, GTPases, phosphatases, heat-shock proteins and DNA unwinding proteins.
The unique formulation of PiColorLock™ affords enhanced assay linearity, dynamic range and colour stability; it is also possible with PiColorLock™ to work with unstable substrates (e.g. ATP, GTP) that give high non-enzymatic background signals with other acidic dye-based detection reagents.
PiColorLock™ - phosphate detection reagent features and benefits
|Colorimetric assay||Simple, scalable and non-radioactive|
|Compatible with almost any assay buffer||Flexibility over a wide range of existing assays|
|Stable reagent formulation||Long shelf life|
|Unique accelerator||Speeds up color development|
|Unique stabilizer||Suppresses non enzymatic backgrounds with acid-labile substrates (ATP or GTP)|
|PiColorLock™-Pi complex is very stable||No precipitation, results can be measured over several hours – ideal for HTS|
|Wide linear range||No inhibition of color development by high concentrations of protein|
The unique PiColorLockTM Stabilizer prevents background drift with time
The PiColorLockTM reagent is often used with unstable substrates (e.g. ATP, GTP) that give rise to non-enzymatic background drift with time. The unique stabilizer, provided in the PiColorLockTM phosphate detection system, blocks this non-enzymatic breakdown generating a stable low background. The stable signal can therefore be read up to several hours after the reaction has ended.
For more information on PiColorLock™ take a look at our Phosphate Detection Guide.
Please do not hesitate to Contact us to find out more!/p>
Figure 1. Competitor assays are beset with several problems including reagent precipitation. The high stability of PiColorLock™ ensures high stability of the colored dye-phosphate complexes (green color).
Figure 2. ATP has been incubated in three detection reagents. A steadily rising background signal is seen with competitor reagents, whereas PiColorLock™ gives baseline readings.
Figure 3. PiColorLock™ has also been designed to have a large linear range, thus reducing the need for sample dilution. For the purposes of comparison, data for PiColorLock™ ALS reagent (no longer available) is also shown, as it provides similar absorbance values to competitor products, at least at low levels of inorganic phosphate. Competitors’ products are linear over a much narrower range of concentrations.
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™ 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™?
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.
Which reagents does the PiColorLock™ Phosphate Detection System contain?
|Reagent||Product Code: 303-0030||Product Code: 303-0125|
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