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

Description
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.
Features
Read-out of results: Quantitative
Detection system: Luminescence
Regulatory status: Research Use Only
Applications
Quantification of functional C5a
Detection of C5aR inhibitory activity
Principle
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.
Size
1 vial corresponding to one 96 well plate
Species Reactivity
Human
Cell Background
HEK293 Cells
Sample
Works with both adherent and suspension cells.
Growth Medium
Gibco (cat no 12648-010)
Cell Purity
>95%
Cell Viability
>90%
Sterility Testing
Creative Biolabs provides sterility testing in accordance with USP and EP regulations. All of our sterility testing is performed in an isolator or clean room environments. The cell line has been screened using the membrane filtration testing methods to confirm the absence of aerobic, anaerobic and fungi microorganisms.
Identity Testing
Identity testing is required for newly established cell lines. Isoenzyme analysis is used to confirm the identity of the species of a cell line. Alternative methods for identity testing include DNA fingerprinting, STR analysis and karyology.
Virological Safety Testing
A broad range of viruses is susceptible to affecting human cell lines. We can provide in vivo/vitro virus saftey assays by utilizing various animal systems. These viruses include: adventitious viruses, bovine viruses, human and simian viruses, porcine viruses, retrovirus and rodent viruses.
Genetic Stability Testing
We perform cell genetic stability studies under ICH guidelines. We can provide guidance on the appropriate testing program upon your requirements.
Mycoplasma Testing
Guaranteed mycoplasma-free
Shipping
Dry ice
Storage
Upon receipt confirm that adequate dry-ice is present, and the cells are
frozen. Immediately transfer to -80°C storage. Cells should be stored at
-80°C (do not store at any other temperature) and are stable as
supplied until the expiry date shown. Cells should be used within 30 min
of thawing.
Warnings
Avoid multiple freeze/thaw cycles
Research Use Only
Our recombinant cell are for research use only, not for diagnostic or therapeutic use.
Quality Control
The expression of hVISTA has been verified by flow cytometry.
The induction of antibody-dependent cellular cytotoxicity (ADCC) has been verified using anti-hVISTA-hIgG1 antibody and Jurkat-NFAT Lucia™ CD16 reporter cell line.
The stability of the 20 channels after thawing has been verified.
Raji-hVISTA cells are guaranteed to be free of mycoplasma.

Figure 1 Example of C5a calibration curve.

All products and services are for Research Use Only. Do Not use in humans.

ONLINE INQUIRY

Creative Biolabs has established a team of customer support scientists ready to discuss ADCC/CDC optimization strategies, antibody production, bioinformatics analysis and other molecular biology/biotechnology issues.

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