Simple, no-wash kit for robust detection of Phoshpo-VEGRF2 (Tyr1175)
HIF-1 alpha Cellular Kit
- High sensitivity
- Faster and more convenient than ELISA
HIF-1a is one of the 2 subunits of the transcription factor hypoxia-inducible factor-1 (HIF-1). HIF-1 acts as a master regulator of cellular and systemic response to hypoxia by activating the translation of genes involved in energy metabolism, angiogenesis, and apoptosis, among others. High levels of HIF-1 have been associated with aggressive tumor progression. HIF-1a is also strongly linked to diabetes through its involvement in the glucose uptake of target cells. It has also been implicated in diabetes complications, such as diabetic nephropathy or diabetic wound repair.
Human MCF-7 epidermoid carcinoma cells were plated at 50,000 cells/well in a 96 well plate. After a preincubation with increasing concentrations of 1mM DFO for 30 minutes to prevent the degradation of HIF-1α , the cells were treated for 6 h at 37°C with increasing concentrations of the PI3K inhibitor LY294002 for 30 minutes (in triplicate) to inhibit the PI3K/AKT pathway. The cells were then activated with 10 nM IGF-1 and 1mM DFO. Next, the medium was removed and the cells were lysed with 50 µL of supplemented lysis buffer#4 for 30 min at RT under gentle shaking. 16 µL of lysate were then transferred into 2 different wells of a 384-sv plate for measurement in parallel.
HIF1 consists of a constitutively expressed HIF-1ß subunit (87 kDa) and an O2-regulated HIF-1a subunit (120 kDa) which contains an O2-dependent degradation domain (ODD).
Under normoxia, HIF-1a is hydroxylated on its ODD domain by O2-activated prolyl hydroxylases (PHDs), leading to its rapid ubiquitination and subsequent degradation by the proteasome.
Under hypoxia, PHD activity is inhibited and HIF-1a accumulates in the cytosol, translocates to the nucleus, heterodimerizes with HIF-1ß, and is activated. The complex activates the transcription of more than 100 target genes that facilitate responses to the hypoxic environment by encoding proteins involved in angiogenesis, erythropoiesis, cell proliferation/survival, and inflammation.
Simple and robust detection kit for Phospho-EGFR (Tyr1068)
Simple and robust detection kit for Pan-Phospho-EGRF
Assay kit for the detection of Phospho-NDRG1 (Thr346)
All-in-one kit for robust detection of Total NDRG2
Simple and robust detection kit for Total EGFR
Assay kit for the detection of Phospho-p53 (Ser15)
All-in-one kit for robust detection of Total p53