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33-017

Sigma-Aldrich

PI 3-Kinase HTRF® Assay; 1920 wells

The PI 3-Kinase HTRF Assay is a high-performance assay kit that provides a universal method for testing all Class I PI3-Kinases in a homogeneous format. This pack size has a screening capacity of 5 plates.

Synonym(s):

Kinase activity assay

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About This Item

UNSPSC Code:
12161503
eCl@ss:
32161000
NACRES:
NA.84

Quality Level

species reactivity

mouse, human

manufacturer/tradename

HTRF®
Upstate®

technique(s)

activity assay: suitable (kinase)

NCBI accession no.

UniProt accession no.

detection method

fluorometric (HTRF)

shipped in

dry ice

Gene Information

human ... PIK3CA(5290)

General description

The PI3-Kinase HTRF Assay is a high-performance assay kit that provides a universal method for assaying all Class I PI3-Kinases in a homogeneous 384-well format.

Application

PI 3-Kinase HTRF® Assay; 1920 wells have been used to measure the activity of phosphoinositide-3-kinases (PI3Ks). It has also been used in a biochemical assay to measure the potency and selectivity of LL-00071210 (a selective PI3Kδ inhibitor).

Biochem/physiol Actions

The PI 3-Kinases catalyze the phosphorylation of the three positions of phosphatidylinositols. The Class I enzymes convert phosphatidylinositol 4,5-bisphosphate (PIP2) to phosphatidylinositol 3,4,5-trisphosphate (PIP3) in the presence of ATP and Mg2+.

Features and Benefits

  • Large Stokes shift and time-resolved elements of technology help to reduce compound interference, minimizing concern of false negatives
  • Picomole sensitivity of the HTRF assay platform enables the integrity of enzyme kinetics to be retained, ensuring the most physiologically relevant assay parameters.
  • Compatible with all Class I phosphoinositide 3-kinase (PI3K) isoforms: enables relative data comparisons and eliminates the need for multiple assay types.
  • Reliable, proven assay—based on the technology used in the Millipore, Upstate® KinaseProfiler Service.
  • Z` factor routinely exceeds 0.8, ensuring consistent, reliable data.

Packaging

Kit capacity: 1920 wells

Components

4X Reaction Buffer

PIP2 (Substrate; 1mM stock)

Stop Reagent A

Stop Reagent B

DM A Detection Reagent

DM B Detection Reagent

DM C Detection Reagent

Legal Information

HTRF is a registered trademark of Cis Bio International
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Pictograms

Exclamation mark

Signal Word

Warning

Hazard Statements

Hazard Classifications

Eye Irrit. 2

Storage Class Code

10 - Combustible liquids

WGK

WGK 2


Certificates of Analysis (COA)

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Robert H Sinnamon et al.
Protein expression and purification, 73(2), 167-176 (2010-05-12)
Phosphoinositide 3-kinases have been targeted for therapeutic research because they are key components of a cell signaling cascade controlling proliferation, growth, and survival. Direct activation of the PI3Kalpha pathway contributes to the development and progression of solid tumors in breast
Reine Nehmé et al.
Analytical and bioanalytical chemistry, 406(15), 3743-3754 (2014-05-13)
Human kinases are one of the most promising targets for cancer therapy. Methods able to measure the effects of drugs on these cell agents remain crucial for biologists and medicinal chemists. The current work therefore sought to develop an in-capillary
Marinela Méndez-Pertuz et al.
Nature communications, 8(1), 1278-1278 (2017-11-04)
Telomeres and the insulin/PI3K pathway are considered hallmarks of aging and cancer. Here, we describe a role for PI3K/AKT in the regulation of TRF1, an essential component of the shelterin complex. PI3K and AKT chemical inhibitors reduce TRF1 telomeric foci

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