Product category: In vitro assays
Product code: KeratinoSens-Assay

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 LineKeratinoSens™ cell line

Assay ProtocolThe 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 Samples10ml of liquids/10g of solids

Application and Test samplesThree independently performed experiments
12 concentrations in triplicate
2000 μM as highest concentration (as per OECD guideline)

Quality ControlsVehicle control: 1% DMSO, medium
Reference control: cinnamic aldehyde

Time Point48 hours

Data AnalysisCell viability and induction of luciferase reporter gene expression

Data DeliveryImax 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 predictionSignificant induction of luciferase activity >1.5 fold
EC1.5 is <1000μM
Cell viability >70%
Dose response for luciferase induction

Vendor: Curio Biotech


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