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Expression Profiling of Clinically Relevant CYPs Assay Kit

SIZE SKU PRICE
2 x-48 assays in-96 well format
SIZE SKU
2 x-48 assays in-96 well format

Product Description and Product Data

This Expression Profiling of Clinically Relevant CYPs assay kit contains optimized reagents for the culturing and treatment of upcyte® hepatocytes to assess drug-induced changes in CYP3A4, CYP1A1, CYP2B6, CYP2C8, CYP2C9, CYP2C19, and CYP2E1.

The kit provides two aliquots of upcyte® hepatocytes, two cell culture-ready assay plates, optimized Cell Culture Medium for use in all steps of the assay procedure, and three reference compounds (rifampicin, β-naphthoflavone, and CDCA) that activate one or more of the primary xenobiotic-sensing receptors: PXR, CAR, AhR, and FXR. Upon activation, these nuclear receptors modulate the expression of the CYP genes. Also included are seven sets of validated qPCR primers for quantifying drug-induced changes in the expression of CYP3A4, CYP1A1, CYP2B6, CYP2C8, CYP2C9, CYP2C19, and CYP2E1, as well as primers for ACTB.

Please note: This kit does not include reagents or protocols for cell lysis, RNA isolation, cDNA preparation, or qPCR assays.

Features

  • Clear, Reproducible Results

  • All-Inclusive Assay Systems
  • Exceptional Cell Viability Post-Thaw
  • Consistent Results Lot to Lot

Product Specifications

SpeciesHuman
Kit Components
  • upcyte® Hepatocytes
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • Rifampicin, (positive control for modulators of CYP3A4 and CYP2C genes; in DMSO)
  • b-Naphthoflavone, (positive control for modulators of CYP1 and CYP2 genes; in DMSO)
  • CDCA, (positive control for modulators of CYP1A1, CYP2C, CYP2E1, and CYP3A4 genes); in DMSO)
  • 8 Primer sets (all prepared in water at 20x concentrations)
  • • CYP3A4
  • • CYP1A1
  • • CYP2B6
  • • CYP2C8
  • • CYP2C9
  • • CYP2C19
  • • CYP2E1
  • • b-Actin (ACTB)
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Shipping RequirementsDry Ice
Storage temperature-80C

Data

Target Background

The reagents and materials provided in the assay kit for the Expression Profiling of Clinically Relevant CYPs are formatted to allow for two alternative cell culture setups. In one scenario 48 culture wells may be set up at two different times. In the other assay scenario 96 culture wells may be set up at one time. The kit includes two aliquots of upcyte® hepatocytes, donor 10-03, isolated from an adult Caucasian female. These hepatocytes are cryopreserved using INDIGO’s proprietary CryoMite™ process, which yields high cell viability post-thaw, and provides the convenience of immediately dispensing cells into assay plates. There is no need for intermediate treatment steps such as spin-and-rinse of cells, viability determinations, or cell titer adjustments prior to assay setup. During an overnight culture period for cell recovery the hepatocytes will form a confluent monolayer that is ready to receive treatment media containing the user’s test compounds.

The kit provides two aliquots of upcyte® hepatocytes, two cell culture-ready assay plates, optimized Cell Culture Medium (CCM) for use in all steps of the assay procedure (cell thawing, seeding, and preparation of treatment media), and three reference compounds (rifampicin, β-naphthoflavone, and chenodeoxycholic acid (CDCA)) that activate one or more of the primary xenobiotic-sensing receptors: PXR, CAR, AhR, and FXR. Upon activation, these nuclear receptors modulate the expression of the clinically relevant CYP genes. Also included are seven sets of validated qPCR primers for quantifying drug-induced changes in the expression of CYP3A4, CYP1A1, CYP2B6, CYP2C8, CYP2C9, CYP2C19, and CYP2E1, as well as primers for ACTB (β-actin; the internal control used to normalize all CYP gene expression data).

Please note: This kit does not include reagents or protocols for cell lysis, RNA isolation, cDNA preparation, or qPCR assays.

Citations

Cranberry contains high levels of nutrients and bioactive molecules that have health‐promoting properties. The purpose of the present studies was to determine if cranberry extracts (CEs) contain phytochemicals that exert anti‐inflammatory effects. The human monocytic cell line THP‐1 was treated with two CEs (CE and 90MX) and subsequently challenged with Lipopolysaccharides (LPS). Tumor necrosis factor α (TNF α) expression was decreased in the CE‐treated cells, indicative of an anti‐inflammatory effect. Gene expression microarrays identified several immune‐related genes that were responsive to CEs including interferon‐induced protein with tetratricopeptide repeats 1 and 3 (IFIT 1 and 3), macrophage scavenger receptor 1 (MSR1) and colony‐stimulating factor 2 (CSF2). In addition, in the CE‐treated cells, metallothionein 1F and other metal‐responsive genes were induced. Taken together, this data indicates that CEs contain bioactive components that have anti‐inflammatory effects and may protect cells from oxidative damage.
2016-04-25

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Expression Profiling of Clinically Relevant CYPs

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