Applied Biosystems™

PureQuant™ CD8+ T-Cell Assay

Catalog number:  A43674

Related applications:

Real Time PCR (qPCR)

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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.

The PureQuant CD8+ T-Cell Assay utilizes the CD8B gene as an epigenetic marker for CD8+ T cells; the assay targets regulatory elements within its third intron (see http://stm.sciencemag.org/content/10/452/eaan3508).

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 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 CD8 Module (CD8 & GAPDH TaqMan primers and probes, Calibrator DNA, Reference DNA, Lambda DNA, Standard DNA), store at -5 to -30°C