Product Name Catalog # Price   Qty
Human HIF-Regulated cDNA Profiling in Cell Lysates AP-3102 $489
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Characterization

Description:

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 CLTM 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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