Human P450 cDNA Plate Array

The cytochrome P450 are a multiple gene family of enzymes, playing central roles in oxidative metabolism of a wide range of xenobiotics, including chemicals, sterols, fatty acids and anticancer drugs. Many of them can activate cancer carcinogen, which contribute to tumorgenesis. Human CYP genes encode fifty-seven P450 proteins with numerous homolog variants in different tissues. They are highly differentially expressed in response to a variety of environmental chemicals and pharmaceuticals. Signosis developed P450 cDNA Plate Array for profiling the expression of a panel of 22 CYP genes.

A. Benefits:

Based on Signosis' proprietary cDNA plate array detection technology, we developed Human P450 cDNA plate array, which offers following benefits:

  • Simple - Do the gene expression profiling as doing ELISA.

  • High sensitive detection- Detected mRNA molecules are converted into multiple biotin-labeled cDNAs for sensitive detection.

  • Quantitative comparison – The difference of two or more samples in gene expression can be quantitatively analyzed.

B. Principle of the technology

Signosis’ proprietary cDNA plate array is a plate-based hybridization profiling analysis for monitoring the expression of dozens of genes through reverse transcription of mRNA into cDNA. Like array analyses, total RNA is first reverse transcribed into cDNA in the presence of biotin-dUTP in the assay. Targeted genes are then specifically captured onto individual wells on a plate, instead of membranes, through a pre-coated gene-specific oligonucleotide. The captured cDNAs are further detected with streptavidin-HRP. The captured cDNAs are further detected with streptavidin-HRP. Luminescence is reported as relative light units (RLUs) on a microplate luminometer. The expression level of genes is directly proportional to the luminescent intensity.

Figure 1
Diagram

Figure 2. Analysis of expression of P450 genes with P450 cDNA Plate Array. 2ug of RNAs from MCF7, HL-60, HeLa and 293 cell lines were subjected to P450 cDNA plate assay. The cDNA plate array were detected with HRP by a luminometer plate reader.

 

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