VEGF Bioassay Technical Manual
Instructions for Use of Product(s)
GA2001, GA2005, GA3001, GA3005, GA4001, GA4005
Literature # TM544
The VEGF Bioassay is a bioluminescent cell-based assay that measures VEGF stimulation and inhibition of KDR (VEGFR2) using the NFAT-RE as a readout. This assay overcomes many of the limitations of the current endothelial cell proliferation assays, and can be used for the discovery and development of novel biologic therapies aimed at either inducing or inhibiting the VEGF response. The KDR/NFAT-RE HEK293 Cells are provided in a thaw-and-use format as cryopreserved cells that can be thawed, plated and used in an assay without the need for cell propagation (also offered as a cell propagation model; CPM, Cat.# GA1082).
The KDR/NFAT-RE HEK293 Cells have been engineered to express the NFAT response element upstream of Luc2P as well as exogenous KDR. When VEGF binds to the KDR/NFAT-RE HEK293 Cells, the KDR transduces intracellular signals resulting in NFAT-RE-mediated luminescence. The bioluminescent signal is detected and quantified using Bio-Glo™ Luciferase Assay System and a standard luminometer, such as the GloMax® Discover System.
This Technical Manual is relevant for Cat.# GA2001 and GA2005; GA3001 and GA3005 (Korea); GA4001 and GA4005 (Taiwan).
Summary of Changes
The following changes were made to the 12/22 revision of this document:
1. In Section 4, Note a., the suggested starting concentration of test and reference samples was changed from 18μg/ml to 6μg/ml.
2. Miscellaneous formatting updates.
3. “Notes” were changed from numerical listings to alphabetical listings.
4. Disclaimers were updated.
5. Kit component storage temperatures were updated to show temperature range.
6. The template, TM cover and font were updated.
Revised 12/22.