AD Model: Beta-Secretase Expression Assay tGFP-BACE1 HEK293 Cell Line
A novel HEK293/tGFP-BACE cell line has been developed through stable transfection for monitoring the expression of beta-secretase in cell-based assays. HEK293/tGFP-BACE cell line was obtained by transfection of an expression vector for a fusion protein of turboGFP and human BACE, as well as G418 into HEK cells with Evrogen TurboGFP. Cells were then grown in the presence G418. Single cells with strong green fluorescence were selected by flow cytometry, and allowed to expand. These cells constitutively express the tGFP-BACE fusion protein.
Assay Details: Recombinant HEK293 cells stably expressing human beta secretase (BACE1) fused to the turbo green fluorescent protein (tGFP) can be use as a beta-Secretase Expression Assay Kit, designed to assay for compounds that inhibit the production of beta-secretase by quantifying the fluorescence by accumulation of tGFP-BACE1 in the cytoplasm. tGFP-BACE can be imaged on most fluorescent microplate readers, HCS platforms or fluorescence microscopes.
Fig 1: Left: HEK293 BACE-tGFP cells culture image by phase contrast microscopy
Middle: Nuclear DAPI staining of HEK293BACE-tGFP cells culture image by epifluorescence microscopy
Right: Fluorescent tGFP-BACE HEK293 cells culture image by epifluorescence microscopy.