PureQuant™ Th17 Assay
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PureQuant™ Th17 Assay
Applied Biosystems™

PureQuant™ Th17 Assay

The Applied Biosystems PureQuant assays are a family of assays that accurately identify and quantify specific immune cell types inRead more
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Catalog number A43676
Price (USD)
4,120.00
Each
Add to cart
Price (USD)
4,120.00
Each
Add to cart
The Applied Biosystems PureQuant assays are a family of assays that accurately identify and quantify specific immune cell types in a mixed population of cells. These assays are TaqMan qPCR-based and measure genomic DNA methylation at specific sites that serve as a unique identifier. The assays include standards and controls and each 96-well plate can accommodate up to seven test samples. Results are provided as a percentage of target cell type out of total input cells using a free analysis tool.

PureQuant Th17 Assay utilizes a specific demethylated site in the pro-inflammatory cytokine interleukin 17A (IL17). This cytokine is exclusively produced by T helper 17 cells.

Benefits include:
Proven performance―core technology has been used for identification and quantification of immune cell types for over a decade
Easily standardized―qPCR method enables consistent results across multiple users, instruments, and sites
Facilitates regulatory filings―established performance specifications following ICHQ2(R1) guidelines
Scheduling flexibility―assay can be performed using fresh or frozen cells/gDNA

The assays include all reagents, standards, and controls required to run the assay. Specific components include:
Calibrator: corrects for variation in bisulfite conversion efficiency. Plasmid-based calibrator harbors the genomic target regions in equimolar ratios and includes GAPDH. Subjected to bisulfite conversion followed by copy number estimation via qPCR standard curve. Allows estimation of assay-specific bisulfite efficiency factor: the ratio between TpG/GAPDH copy numbers measured from the same plasmid.
Reference: checks assay performance. Consists of genomic DNA extracted from human blood of pooled healthy donors. The reference values represent the expected range of the target cell type. For example, CD8+ cells are generally more abundant than Treg or Th17 cells.
Standards: determine the unknown copy number. Consist of six dilutions spanning 4-log linear dynamic range (30-31250 copies). Absolute copy numbers can reliably be determined using the slope and intercept of the respective standard curves.

This product is manufactured at a site with an ISO9001-certified quality management system.

For Research Use or Manufacturing of Cell, Gene, or Tissue-Based Products. CAUTION: Not intended for direct administration into humans or animals.
Specifications
Detection MethodPrimer-Probe Detection
For Use With (Application)Cells, DNA
Quantity28 reactions
Shipping ConditionApproved for shipment at Room Temperature or on Wet or Dry Ice
Unit SizeEach
Contents & Storage
  • Dynabeads SILANE Genomic DNA Kit, store at 2°C to 8°CC
  • 1 x 2 mL PureQuant qPCR Master Mix, store at 2°C to 8°CC
  • 4 x ABS
  • THFA, store at room temperature
  • PureQuant Th17 Module, store at -15 to -25°C (CpG & TpG TaqMan primers and probes, Reference DNA, Lambda DNA, Standard DNA)
  • PureQuant Hot Start PCR
  • PureQuant Pre-amplification primer

Frequently asked questions (FAQs)

In the PureQuant assays, can I make changes to the data analysis template?

No, the template is locked and no changes can be made to the formulas and calculations. In the event of a bad/outlier replicate (or replicates), consult the instruction tab of the template for guidance on performing analysis in these cases.

If I am new to the PureQuant technology, do you have a suitable product that I can use as an orthogonal method for comparison?

Yes, we've developed the corresponding eBioscience Essential Phenotyping kits for flow cytometry characterization of T cells, regulatory T cells, and type 1 and type 17 T helper cells. These kits were used during the development of the PureQuant CD8+, Treg, and Th17 Assays.

Can alternative bisulfite conversion kits or reagents (for example sodium bisulfite) be used for bisulfite conversion?

No, the bisulfite conversation kit was optimized specifically for the PureQuant Assays. The ammonium bisulfite and THFA were selected from specific suppliers and using alternatives will jeopardize performance of the assays.

Do the PureQuant assays require specific qPCR instrumentation to run?

No. The assays have been demonstrated to work on Thermo Fisher qPCR instrumentation and other competitor platforms. The only requirement is that the instrument must be compatible with FAM and non-fluorescent quencher or their respective equivalents.

Since the PureQuant assay technology is licensed-in from a third party, are there any restrictions on its use?

Yes, the Limited Use Label License is as follows:
Notice to Purchaser: The purchase of this product conveys to the purchaser the limited, non-transferable rights under licensed patents for the purchaser to use the product only to (a) perform research and development intended to support the manufacturing of cell therapy products that are intended for subsequent administration to human patients and (b) quality control testing for use in the manufacture of cellular therapy products that are intended for subsequent administration to human patients. This product is not for use in any testing or analysis for any other purpose, any provision of services or any third party commercial use conducting clinical trials monitoring services on patient samples. No right to resell this product or any of its components is conveyed expressly, by implication, or by estoppel. For information on obtaining additional rights, please contact outlicensing@thermofisher.com, Licensing and Commercial Supply, Thermo Fisher Scientific, 5823 Newton Drive, Carlsbad, CA, 92008, United States