This non radioactive colorimetric assay kit contains all the necessary reagents for measuring enzyme activity (everything included in the PiColorlock kit and more) and is ideal for high throughput drug screening.
The kits contain our PiColorlock, a non-radioactive, superior phosphate detection reagent.
Key Features of PiColorLock:
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.
Colorimetric assays for GTPases are invariably based on the formation of colored complexes between an inorganic phosphate and a dye molecule under acidic conditions.
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FAQs for GTPase Kit (602-0120). (Showing 1 - 7 of 7 FAQs).
May we have the protocol for 602-0120?The info. from http://www.novusbio.com/support/protocols/colorimetric-gtpase-assay-kit-protocol.html seems insufficient.
Here is the corresponding full protocol for your customer for this kit. I hope they find this information useful. http://www.innovabiosciences.com/images/stories/innova/pdfs/gtpase_assay_system_protocol_r3.pdf
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.
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 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 stabilizer with the sample and detection reagent. Make sure the stabilizer 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.
This is regarding the GTPase assay kit catalog No: 602-0120. I want to know what is the equipment one needs to take the reading as kit contents are pipetted out in a 96-well plate. Does this require a fluorescence plate reader or will it be okay to take reading on the spectrophotometer if a cuvette is used.
It is possible to carry out the detection step using cuvettes, but sufficient volume for the cuvette must be used, and note that the PiColorLock solution may damage the cuvette.
At which temperature does the assay have to be performed?
The assay can be carried out at room temperature, as the kit contains proprietary buffers which reduce background hydrolysis (the assay is less temperature sensitive because of this). Please note that it is important for all reagents to be warmed up to room temperature before the assay is carried out.