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
• 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:
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.
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.
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.