CDC C5a (+) Assay Cell Line, Human HEK293 Cells (CAT#: AD-P023-C) Datasheet

The iCrea™ C5a Assay Cell Line can be used for the quantification of C5a activity, and for determination of inhibitory activity against either C5a itself or against the C5a receptor in test samples, including human serum. iCrea™ C5a Assay Cell Line are human HEK293[1] cells ( CRL-1573) which have been genetically engineered and optimized to be responsive to human complement component 5a (C5a), resulting in a proportional expression of Firefly Luciferase. Normalization of cell counts, and serum matrix effects is obtained by a second reporter gene, a Renilla Luciferase reporter gene construct, under control of a constitutive promotor.

Read-out of results: Quantitative
Detection system: Luminescence
Regulatory status: Research Use Only

Quantification of functional C5a
Detection of C5aR inhibitory activity

The iCrea™ C5a Assay Cell Line are engineered cells, optimized to express Firefly luciferase (FL) under the control of a C5a responsive promoter and exhibits a dose-dependent increase in firefly luciferase (FL) reporter gene activity following treatment of cells with increasing concentrations of human C5a. (Fig.1). The cells also contain the Renilla luciferase (RL) internal standardization gene under the control of a constitutive promoter that renders assay results independent of cell number and provides a means for correcting for cytotoxic effects that may be encountered with some biological samples. Some increase in RL reporter-gene activity may be observed, however, at high non-physiologic concentrations of C5a and consequently normalization of C5a-induced FL activity relative to the expression of RL activity is not recommended for routine use. Normalization of C5a-induced FL activity relative to the expression of RL activity remains, however, a valuable means of correcting for sample cytotoxicity. The luciferase signals can be measured in a luminometer following addition and incubation of luciferase substrate.