Thermo Scientific™ LanthaScreen™ IGF-1R GripTite™ Cell Line

Description

• Description The IGF-1R/PI3K/AKT pathway mediates signals for cell growth, cell survival, transcription, translation, and glucose uptake
• IGF-1R is a receptor tyrosine kinase (RTK) that resides at the top of the pathway
• Upon growth factor binding extracellularly, this RTK undergoes auto-phosphorylation at multiple intracellular tyrosine residues (resulting in pathway activation)
• The LanthaScreen™ IGF-1R GripTite™ (HEK 293 MSR) cellular assay utilizes a human cell line that constitutively expresses IGF-1R fusion proteins with yellow fluorescent protein (YFP)
• This kinase target was introduced using lipid transfection and these cells are a clonal population isolated by FACS, using YFP 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 multiple tyrosine residues on IGF-1R is detected in cell lysates using a generic terbium-labeled phosphospecific antibody (Tb-anti-pY20)
• 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.