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KeratinoSens™
KeratinoSens™ Assay
Turn Around time: 6-8 weeks
The KeratinoSens™ cell line addresses Key Event 2 (KE 2) that occurs in the epidermal keratinocyte gene expression associated with specific cell signalling pathways. The KeratinoSens™ cell line is a cell line derived from human keratinocytes that contains a stably transfected luciferase gene under the transcriptional control of a constitutive promoter fused with an ARE (an antioxidant/electrophile response element). Once a sensitizer binds to the ARE, the luciferase gene is upregulated.
Upregulated luciferase activity is measured using a standard substrate. Based on the results, the test chemical is classified as a Positive sensitizer if the activity is more than or equivalent to 1.5 fold induction or as Non-Sensitizer if it is less than 1.5 fold induction.
| Cell Line | KeratinoSens™ cell line |
| Assay Protocol | The KeratinoSens™ cell line is seeded and grown for 1 day either in Fetal Bovine Serum or Xeno-free (human serum) based cell culture medium.Treatments with the test compounds for 48 hours, then cell viability is evaluated using Resazurin and luciferase reporter gene expression through ONE-Glo Luciferase Assay System. |
| Required Test Samples | 10ml of liquids/10g of solids |
| Application and Test samples | Three independently performed experiments 12 concentrations in triplicate 2000 μM as highest concentration (as per OECD guideline) |
| Quality Controls | Vehicle control: 1% DMSO, medium Reference control: cinnamic aldehyde |
| Time Point | 48 hours |
| Data Analysis | Cell viability and induction of luciferase reporter gene expression |
| Data Delivery | Imax Maximal fold induction (maximum response) EC1.5 value of luciferase activity IC50 values of cell viability Dose response curves for luciferase induction and cell viability |
| Sensitizer prediction | Significant induction of luciferase activity >1.5 fold EC1.5 is <1000μM Cell viability >70% Dose response for luciferase induction |
Vendor: Curio Biotech
