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OCT4-RT-PCR real time

Quantification assay of stemness markers OCT4          cod. BM-009

Principle of the test: Quantitative analysis of OCT4 mRNA expression

Technology: Relative Quantitative Real Time PCR

Gene Target: OCT4

Specimen: cDNA

Results within: ΔΔCt method

Reporting Units: Arbitrary Units (AU)

Number of tests: 25 tests BM-009 

Kit storage: -20°C

Necessary equipment: Thermocycler, 7500 Real Time PCR System

Status: Ready to use

OCT4-RT-PCR real time cod. BM-009​

Quantification assay of stemness markers OCT4

·      OCT4-RT-PCR real time Quantification complete kit 25 tests BM-009

Oct-4, a transcription factor also known as Oct-3, Oct-3/4, Otf3 or NF-A3, is encoded by the Pou5f1 gene (located on chromosome 6 in human and 17 in mouse) and belongs to the POU (Pit, Oct, Unc) family of DNA binding-proteins. These proteins regulate the expression of target genes by binding to the octamer motif ATGCAAAT within their promoter or enhancer regions16,17. Oct4, whose expression is associated with pluripotent properties of stem cells, is an essential factor controlling early stages of mammalian embryogenesis18-20.

We have established a novel quantitative analysis of OCT4 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 OCT4-RT-PCR real time Quantification kit is suitable for any laboratory.

Reference

1.      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

2.      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.

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

4.      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

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