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PIP3 Quantification TR-FRET Assay
Description:
PIP3 Quantification TR-FRET Assay
Qty/Pk:
1 plate
Product Overview:
PI 3-Kinases α, β, γ and δ (h) catalyse the phosphorylation of phosphatidylinositol 4,5-bisphosphate (PIP2) to phosphatidylinositol 3,4,5-trisphosphate (PIP3) in the presence of ATP and Mg2+. The PIP3 product is detected by displacement of biotin-PIP3 from an energy transfer complex consisting of Europium labelled anti-GST monoclonal antibody, a GSTtagged pleckstrin homology (PH) domain, biotinylated PIP3 and Streptavidin-Allophycocyanin (APC). Excitation of Europium in the complex results in an energy transfer to the APC and a fluorescent emission at 665 nm. The PIP3 product isolated from cell membranes displaces biotin-PIP3 from the complex resulting in a loss of energy transfer and thus a decrease in signal.
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Applications:
The PIP3 Quantification TR-FRET Assay kit provides an easy to use method for assaying PIP3 levels by detecting the displacement of biotin-PIP3 from an energy transfer complex.
Key Applications:
Kinase Assay
Application Notes:
This assay requires lipid extraction from cells. A protocol is included with this assay.
Usage Statement:
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.
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Entrez Gene Summary:
Phosphatidylinositol 3-kinase is composed of an 85 kDa regulatory subunit and a 110 kDa catalytic subunit. The protein encoded by this gene represents the catalytic subunit, which uses ATP to phosphorylate PtdIns, PtdIns4P and PtdIns(4,5)P2. This gene has been found to be oncogenic and has been implicated in cervical cancers.
UniProt Summary:
FUNCTION: SwissProt: P42336 # Phosphorylates PtdIns, PtdIns4P and PtdIns(4,5)P2 with a preference for PtdIns(4,5)P2.
SIZE: 1068 amino acids; 124284 Da
SUBUNIT: Heterodimer of a p110 (catalytic) and a p85 (regulatory) subunit. Binds to IRS1 in nuclear extracts. Interacts with RUFY3 (By similarity).
DISEASE: SwissProt: P42336 # Defects in PIK3CA are associated with colorectal cancer (CRC) [MIM:114500]. & Defects in PIK3CA are associated with breast cancer [MIM:114480]. & Defects in PIK3CA are associated with epithelial ovarian cancer (EOC) [MIM:604370].
SIMILARITY: SwissProt: P42336 ## Belongs to the PI3/PI4-kinase family. & Contains 1 C2 domain. & Contains 1 PI3K/PI4K domain.
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Gene Symbol:
p110-alpha
MGC142161
PI3K
MGC142163
PIK3CA
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Product Name:
PIP3 Quantification TR-FRET Assay
Analytes Available:
PI 3-Kinase
Components:
Black 96-well plate
Detection Buffer
DM B
DM A
Stop B
PIP3 Standard
Lipid Recovery Buffer
1M DTT
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UniProt Number:
P42336
Detection Methods:
TR-Fret
HTRF
Packaging:
Kit capacity: 96-wells
Entrez Gene Number:
NM_006218.2 |