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201904151249411

MULTISTEM cod. BM-002 - Multiplex One Step RT-PCR Real Time            Cod. BM-002

 

Principle of the test : Quantitative analysis of stemness genes

Technology : Multiplex One Step RT- PCR Real Time

Gene Target : NANOG OCT4 SOX2

Internal Control: GAPDH

Specimen : RNA

Results : ΔΔCt method  

Reporting Units : Arbitrary Units (AU)

Number of tests : 25 tests BM-002

Kit storage : -20°C

Necessary equipment : 7500 Real Time PCR System or CFX96 DetectionSystem

MULTISTEM Multiplex One Step RT-PCR Real Time cod. BioMole-002 

 

Quantification assay of stemness markers NANOG OCT4 SOX2

 

In human and mouse embryonic stem cells (ESCs), Oct4, Sox2 and Nanog form a core transcriptional regulatory circuitry in pluripotent stem cells to maintain their self-renewal. Oct4 and Sox2 co-occupy a large number of enhancers/promoters and regulate the expression levels of their target genes.

These transcription factors 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, OCT4 and SOX2 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 MULTISTEM Multiplex One step RT-PCR real time kit is suitable for any laboratory.

 

 

 

Reference

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

3.      Scholer HR, Balling R, Hatzopoulos AK, Suzuki N, Gruss P. Octamer binding proteins confer transcriptional activity in early mouse embryogenesis. EMBO J. 1989;8:2551–7

4.      Yeom YI, Ha HS, Balling R, Scholer HR, Artzt K. Structure, expression and chromosomal location of the Oct-4 gene. Mech Dev. 1991;35:171–9.

5.      Boiani M, Scholer HR. Regulatory networks in embryo-derived pluripotent stem cells. Nat Rev Mol Cell Biol. 2005;6:872–84

6.      Nichols J, Zevnik B, Anastassiadis K, Niwa H, Klewe-Nebenius D, Chambers I, Schöler H, Smith A. Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell. 1998;95:379–91

7.      Scholer HR. Octamania: the POU factors in murine development. Trends Genet. 1991;7:323–9.

8.      Zhang S, Cui W. Sox2, a key factor in the regulation of pluripotency and neural differentiation. World J Stem Cells. 2014 Jul 26;6(3):305-11. doi: 10.4252/wjsc.v6.i3.305