Thermo Scientific™

LanthaScreen™ PDCD4 HEK 293E Cell Line

The PI3K/AKT/mTOR pathway mediates signals for cell growth and survival, transcription and translation, and regulated cell death. The significance of자세히 알아보기

The PI3K/AKT/mTOR pathway mediates signals for cell growth and survival, transcription and translation, and regulated cell death. The significance of mTOR has been underscored recently by the identification of two distinct cellular complexes: mTORC1 (includes raptor and is rapamycin sensitive) and mTORC2 (includes rictor and is insensitive to rapamycin). Activation of the pathway by insulin (or other mitogens) leads to the phosphorylation of programmed cell death protein 4 (PDCD4) via mTORC1 (indirectly through the ribosomal protein p70 S6 kinase). LanthaScreen™ PDCD4 HEK293E is a human cell line which constitutively expresses GFP-PDCD4 fusion proteins. This kinase target was introduced using lipid transfection followed by selection with Blasticidin. These cells are a clonal population isolated by FACS using GFP fluorescence as a sorting marker. A homogenous immunoassay (addition only, no wash steps) was developed in 384-well format using a TR FRET-based detection method to monitor the phosphorylation of Ser457 on GFP-PDCD4 using a terbium-labeled phosphospecific antibody. The assay has been optimized for performance under numerous experimental conditions (e.g., cell plating density, DMSO tolerance, insulin stimulation time, etc.) and displays excellent statistical data (Z` > 0.6). Moreover, this cell line has been validated with different stimuli and known inhibitors (shows correct EC50 and IC50 values) and serves as robust cellular readout for mTORC1 signaling.

사양

대체 작용제Insulin

길항제PMA, IGF-1

어세이 입력Cell-based terbium phospho-specific activity (stable)

셀 상태Dividing Cells

유전자 기호PDCD4

일차 작용제Insulin

제품라인LanthaScreen™

수량1 Vial

판독End Point

리포터 유전자GFP (EmGFP)

타겟 입력PDCD4

System TypeLanthaScreen™

Druggable TargetSignaling Pathway

Unit SizeEach