GTPase Colorimetric Assay Kit

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Product Details

Summary
Kit Type
Colorimetric Assay Kit

Order Details

GTPase Colorimetric Assay Kit Summary

Description
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.
Kit Type
Colorimetric Assay Kit

Applications/Dilutions

Publications
Read Publications using
602-0120 in the following applications:

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 26143143)

Packaging, Storage & Formulations

Storage
Storage of components varies. See protocol for specific instructions.

Kit Components

Components
  1. 4 vials of lyophilized ATP (10 vials)
  2. 2 x 96-well plates (5 plates)
  3. 1 x 5 ml of 0.1mM Pi standard (1 x 10 ml)
  4. 1 x 5 ml of Stabiliser (1 x 10 ml)
  5. 1 x 5 ml of 0.5M Tris pH 7.5 (1 x 10 ml)
  6. 1 x 0.25 ml of Accelerator (1 x 0.5 ml)
  7. 1 x 1.5 ml of 0.1M MgCl2 (2 x 1.5 ml)
  8. 1 x 10 ml of PiColorLock (1 x 25 ml)

Notes

This prodcut is manufacture by Innova Biosciences.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.

Publications for GTPase Kit (602-0120)(32)

We have publications tested in 2 confirmed species: Mouse, Fish.

We have publications tested in 1 application: Func.


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Func
(1)
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Fish
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Showing Publications 1 - 10 of 32. Show All 32 Publications.
Publications using 602-0120 Applications Species
Lin SJ, Chiang MC, Shih HY et al. Regulator of G protein signaling 2 (Rgs2) regulates neural crest development through Ppard-Sox10 cascade. Biochim. Biophys. Acta. Dec 13 2016 [PMID: 27979767] (Func, Fish) Func Fish
Franco A, Kitsis RN, Fleischer JA et al. Correcting mitochondrial fusion by manipulating mitofusin conformations. Nature. 2016 Dec 01 [PMID: 27775718]
Hu Y, Keniry M, Palmer SO, Bullard JM. Discovery and Analysis of Natural Product Compounds Inhibiting Protein Synthesis in Pseudomonas aeruginosa. Antimicrob. Agents Chemother. May 31 2016 [PMID: 27246774]
Yin X, Manczak M, Reddy PH. Mitochondria-targeted Molecules MitoQ and SS31 Reduce Mutant Huntingtin-induced Mitochondrial Toxicity and Synaptic Damage in Huntington's Disease. Hum. Mol. Genet. 2016 Feb 16 [PMID: 26908605]
Cherubini M, Puigdellivol M, Alberch J, Gines S et al. Cdk5-mediated mitochondrial fission: A key player in dopaminergic toxicity in Huntington's disease Biochim. Biophys. Acta. 2015 Jul 02 [PMID: 26143143] (Mouse) Mouse
Yan J, Liu XH, Han MZ et al. Blockage of GSK3 beta-mediated Drp1 phosphorylation provides neuroprotection in neuronal and mouse models of Alzheimer’s disease. Neurobiol Aging 2014 [PMID: 25192600]
Ellinger D, Glockner A, Koch J et al. Interaction of the Arabidopsis GTPase RabA4c with Its Effector PMR4 Results in Complete Penetration Resistance to Powdery Mildew. Plant Cell 2014 [PMID: 25056861]
Gundner AL, Hahn I, Sendscheid O et al. The PIKE Homolog Centaurin gamma Regulates Developmental Timing in Drosophila. PLoS One 2014 [PMID: 24845618]
Santhosh KT, Sikarwar AS, Hinton M et al. Thromboxane Receptor Hyper-Responsiveness in Hypoxic Pulmonary Hypertension Requires Serine 324. Br J Pharmacol. 2013 [PMID: 24490858]
Alaganan A, Fentress SJ, Tang K et al. Toxoplasma GRA7 effector increases turnover of immunity-related GTPases and contributes to acute virulence in the mouse. PNAS 2013 [PMID: 24390541]
Show All 32 Publications.

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FAQs for GTPase Kit (602-0120). (Showing 1 - 7 of 7 FAQs).

  1. 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
  2. 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.
  3. 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.
  4. 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.
  5. 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.
  6. 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.
  7. 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.

Additional GTPase Products

GTPase 602-0120

Blogs on GTPase.

Understanding OPA1 and Mitochondrial Function
OPA1 belongs to the Dynamin large GTPase protein family. OPA1 exists as a single-pass membrane protein localized in the mitochondrial inner membrane and also as a soluble form in the mitochondrial intermembrane space. There, it is a key player in fusi...  Read full blog post.

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