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NANOG-RT-PCR real time

Quantification assay of stemness markers NANOG          cod. BM-008

Principle of the test: Quantitative analysis of NANOG mRNA expression

Technology: Relative Quantitative Real Time PCR

Gene Target: NANOG

Specimen: cDNA

Results within: ΔΔCt method

Reporting Units: Arbitrary Units (AU)

Number of tests: 25 tests BM-008

Kit storage: -20°C

Necessary equipment: Thermocycler, 7500 Real Time PCR System

Status: Ready to use

NANOG-RT-PCR real time cod. BM-008​

Quantification assay of stemness markers NANOG

·      NANOG-RT-PCR real time Quantification complete kit 25 tests BM-008

Nanog is a homeobox-containing transcription factor with an essential function in maintaining the pluripotent cells of the inner cell mass and in the derivation of embryonic stem cells (ESCs) from these14. Furthermore, overexpression of Nanog is capable of maintaining the pluripotency and self-renewing characteristics of ESCs under what normally would be differentiation inducing culture conditions. There are currently approximately 25 transcriptional factors (TFs) that have been reported to be expressed in stem cells. Of them, OCT4, SOX2, KLF4, Nanog, and SALL4 comprise a core regulatory network for embryonic stem cell maintenance and self-renewal15. These TFs are highly expressed in embryonic stem cells; in contrast, they are mainly silenced in normal somatic cells, except in small groups of adult stem cell populations.

We have established a novel quantitative analysis of NANOG mRNA expression. This method is based on real-time PCR. The expression levels of the mRNAs were determined from the threshold cycle (Ct), and the relative expression levels were calculated using the 2-ΔΔCt method. For mRNA quantification, the Ct values were normalized to the expression of the GAPDH mRNA level. Results are expressed in corresponding arbitrary units (AU) User friendly and complete, the NANOG-RT-PCR real time Quantification kit is suitable for any laboratory.

Reference

1.      Mitsui, K., Tokuzawa, Y., Itoh, H., Segawa, K., Murakami, M., Takahashi, K., Maruyama, M., Maeda, M., and Yamanaka, S. (2003) Cell 113, 631–642

2.      Zhao W, et al. Embryonic stem cell markers. Molecules. 2012;17(6):6196–236.