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Thermo Scientific™

LanthaScreen™ Akt HEK 293E Cell Line

Catalog number: K1615
Thermo Scientific™

LanthaScreen™ Akt HEK 293E Cell Line

Catalog number: K1615
Catalog Number
K1615
Unit Size
1 vial
Price (USD)
8,140.00
Your Price:
Availability
***
Quantity
Catalog NumberUnit SizePrice (USD)AvailabilityQuantity
K16151 vial
8,140.00
Your Price:
***
Product Overview
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LanthaScreen® GFP Cellular Assays allow for the analysis of post-translational modifications for a number of target proteins in an live-cell context. The PI3K⁄AKT pathway mediates signals for cell growth, cell survival, transcription, translation, and glucose uptake. Because of the complexity of this signaling cascade, especially as applied to the regulation of the mammalian target of rapamycin (mTOR), cell-based methods are critical for proper identification of small-molecule mediators of this pathway. The significance of mTOR as a kinase has been underscored recently by the identification of two distinct multimeric complexes inside the cell: mTORC1 (includes raptor and is rapamycin sensitive) and mTORC2 (includes rictor and is insensitive to acute rapamycin exposure). mTORC2 has been shown to phosphorylate and AKT at residue Ser473 for complete activation of this prosurvival kinase. LanthaScreenTM AKT HEK293E is a human cell line which constitutively expresses GFP-AKT fusion proteins. This kinase target was introduced using lipid transfection and these cells are a clonal population isolated by FACS, using GFP fluorescence as a sorting marker and Blasticidin to maintain cells under selection. Using this cell line, a homogenous immunoassay was developed with a time-resolved FRET (TR-FRET) readout in which the insulin-induced phosphorylation of Ser473 on GFP-AKT is detected in cell lysates using a terbium-labeled phosphor-specific antibody. This cell line has been validated with different stimuli⁄inhibitors and shows correct EC50 ⁄ IC50 values. Moreover, this assay has been optimized for performance under variable experimental conditions (including cell plating density, agonist stimulation time, DMSO tolerance and assay development time) and displays excellent statistical data (Z' > 0.6) and good signal-to-background.

Specifications

Alternate Agonists
Insulin
Antagonists
LY294002, PI-103
Assay Entry
Cell-based terbium phospho-specific activity (stable)
Cell State
Dividing Cells
Detection Method
Fluorescent
Gene Symbol
AKT1
Optimal Response
IGF-1 (EC50 = 0.42 µg/ml)
Primary Agonist
IGF-1
Product Line
LanthaScreen™
Readout
TR-FRET
Reporter Gene
GFP (EmGFP)
Target Entry
AKT1
Cell Line
293E (HEK)
System Type
LanthaScreen™
Druggable Target
Signaling Pathway

Contents & Storage

LanthaScreen™ AKT HEK 293E cells are shipped on dry ice. Store in liquid nitrogen immediately upon receipt, or thaw for immediate use.

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