Applied Biosystems™

PureQuant™ Th17 Assay

Catalog number:  A43676

Related applications:

PCR & Real-Time PCR

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Description

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―regulatory support file provided upon request; 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 Only. Not for use in diagnostic procedures.||Not for uses other than those specified in the Limited Use Label License.

Specifications

Detection Method: Primer-Probe Detection
No. of Reactions: 28
PCR Method: qPCR
Reaction Speed: Fast or Standard
Sample Type: Cells, DNA
Shipping Condition: Approved for shipment at Room Temperature or on Wet or Dry Ice
For Use With (Equipment): Nearly any qPCR instrument, QuantStudio™

Contents & storage

• Dynabeads SILANE Genomic DNA Kit, store at 2–8°C
• 2 x PureQuant Master Mix, store at 2–8°C
• 4 x ABS
• THFA, store at room temp
• PureQuant Treg Module (CpG & TpG TaqMan primers and probes, Reference DNA, Lambda DNA, Standard DNA)
• PureQuant Hot Start PCR
• PureQuant Pre-amplification primer