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The LanthaScreen™IkB alpha GripTite® cell line contains a fusion protein consisting of the cDNA encoding for GFP and IkB (GFP is fused to the N-terminus of IkB alpha) under the control of a CMV promoter stably transfected into GripTite® cells. GripTite® is a modified HEK293 cell line, which is a human kidney cell line which shows inducible activation of the NFkB signaling pathway by a number of different ligands, such as TNF and IL-1. Activation of the NFkB pathway involves the activation of IKK resulting in the phosphorylation of its target protein IkB alpha. This LanthaScreen cell line allows therefore the analysis of IKK activity using GFP-IkB alpha phosphorylation as readout. The GFP-IkB alpha construct was transfected into GripTite® cells using Lipofectamine™2000, followed by selection with Blasticidin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as sorting marker. This cell line is validated for EC50 and Z' under optimized conditions using TNF as ligand for IKK mediated GFP-IkB alpha phosphorylation or ubiquitination. This cell line has also been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance and assay development time. Additional information using alternate stimuli is also available. Academic and non-profit customers, please inquire for special pricing.
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