Product Name Catalog # Price   Qty
Human HIF-Regulated cDNA Plate Array AP-0111 $479
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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). 

Signosis has developed a plate-based array for profiling 20+ HIF-regulated genes. By using this assay, researchers are able to compare gene expression in up to three samples on a single microtiter plate.




Applicable Grid:

List of Applicable Genes

  1 2 3 4 5 6 7 8 9 10 11 12
A BCL2 HIF-2a PDGF-A BCL2 HIF-2a PDGF-A BCL2 HIF-2a PDGF-A BCL2 HIF-2a PDGF-A
B Beta-actin HPRT PDK1 Beta-actin HPRT PDK1 Beta-actin HPRT PDK1 Beta-actin HPRT PDK1
C CA9 IGF-II NDRG1 CA9 IGF-II NDRG1 CA9 IGF-II NDRG1 CA9 IGF-II NDRG1
D CTSD  Leptin SAG CTSD  Leptin SAG CTSD  Leptin SAG CTSD  Leptin SAG
E BHLHB2 MMP1 SLC6A8 BHLHB2 MMP1 SLC6A8 BHLHB2 MMP1 SLC6A8 BHLHB2 MMP1 SLC6A8
F Glut-1 MXI-1 VEGF Glut-1 MXI-1 VEGF Glut-1 MXI-1 VEGF Glut-1 MXI-1 VEGF
G Glut-3 NDRG2 VHL Glut-3 NDRG2 VHL Glut-3 NDRG2 VHL Glut-3 NDRG2 VHL
H HIF-1a PAI-1 Blank HIF-1a PAI-1 Blank HIF-1a PAI-1 Blank HIF-1a PAI-1 Blank

Principle

 

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. Luminescence is reported as relative light units (RLUs) on a microplate luminometer. The expression level of genes is directly proportional to the luminescent intensity.

Data


Human HIF-regulated cDNA plate array analysis.  HeLa cells were treated with and without 200 uM cobalt chloride for 24 hours, RNAs prepared, cDNA synthesized with biotin label and subjected to cDNA plate array hybridization and detection.

Literature

View user manual

Citations

1.siRNA-mediated knockdown of aryl hydrocarbon receptor nuclear translocator 2 affects hypoxia-inducible factor-1 regulatory signaling and metabolism in human breast cancer cells. Edited by Judit Ovádi. Qin XY, Wei F, Yoshinaga J, Yonemoto J, Tanokura M, Sone H.  FEBS Lett. 2011 Oct 20;585(20):3310-5.
 
2.Tasquinimod is an allosteric modulator of HDAC4 survival signaling within the compromised cancer microenvironment. Isaacs JT, Antony L, Dalrymple S, Brennen WN, Gerber S, Hammers H, Wissing MD, Kachhap SK, Luo J, Xing L,Bjork P, Olsson A, Björk A, Leanderson T. Cancer Res. 2012 Nov 13.
 
3.HIF-stabilizing bioactive glasses for directing MSC behaviour . MM Azevedo, O Tsigkou, R Nair, G Jell, J Jones… - Tissue Engineering, 2014