Characterization

Description(s):

Rapidly growing tumors result in hypoxic regions. Adaptive responses of most cells to hypoxia are (1) to produce VEGF and other hypoxia-induced angiogenic cytokines that promote increased tissue vascularization, thereby increasing tissue oxygenation, and (2) to switch metabolically from oxidative phosphorylation to anaerobic glycolysis. Hypoxia-inducible factor 1 (HIF-1) is an oxygen-regulated transcriptional activator that plays essential roles in the process. The HIF-1alpha subunit is oxygen-dependent ubiquitination and proteasomal degradation. In addition, cytokines including interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha) stimulate HIF-1 dependent gene expression. HIF-1 increases the expression of several genes that promote blood flow and inflammation, such as vascular endothelial growth factor (VEGF). To get a better overview of gene expression of HIF-1 regulated genes, Signosis has developed a cDNA Plate Array which couples its CL^TM cDNA synthesis kit which allows reverse transcription directly from cell lysates without RNA preparation. This plate-based assays allows profiling the expression of 20+ HIF-1-regulated genes directly in cell lysates. This assay provides quantitative data, allowing researchers to screen multiple samples, as well as high sensitivity, allowing researchers to use less sample. 

Applicable Grid:

List of Applicable Genes

BCL2	HIF-2a	PDGF-A	BCL2	HIF-2a	PDGF-A	BCL2	HIF-2a	PDGF-A	BCL2	HIF-2a	PDGF-A
Beta-actin	HPRT	PDK1	Beta-actin	HPRT	PDK1	Beta-actin	HPRT	PDK1	Beta-actin	HPRT	PDK1
CA9	IGF-II	NDRG1	CA9	IGF-II	NDRG1	CA9	IGF-II	NDRG1	CA9	IGF-II	NDRG1
CTSD	Leptin	SAG	CTSD	Leptin	SAG	CTSD	Leptin	SAG	CTSD	Leptin	SAG
BHLHB2	MMP1	SLC6A8	BHLHB2	MMP1	SLC6A8	BHLHB2	MMP1	SLC6A8	BHLHB2	MMP1	SLC6A8
Glut-1	MXI-1	VEGF	Glut-1	MXI-1	VEGF	Glut-1	MXI-1	VEGF	Glut-1	MXI-1	VEGF
Glut-3	NDRG2	VHL	Glut-3	NDRG2	VHL	Glut-3	NDRG2	VHL	Glut-3	NDRG2	VHL
HIF-1a	PAI-1	Blank	HIF-1a	PAI-1	Blank	HIF-1a	PAI-1	Blank	HIF-1a	PAI-1	Blank

Principle

Unlike conventional reverse transcription which needs total RNA, Signosis has developed a CL(TM) cDNA synthesis-based plate array. It can synthesize cDNA probes through reverse transcription directly in cell lysates without RNA preparation. The synthesized cDNA probes are then applied for a plate array hybridization. Targeted genes are specifically captured onto individual wells on a plate through a pre-coated gene-specific oligonucleotide. 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.

 

 

Literature

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