IOX 2, inhibitor

IOX 2

Cat. Num. AAA575732

• Purity: 98.0%
• Synonyms: IOX 2; inhibitor

• Purity/Purification: 98.0%
• Form/Format: POWDER
• Sequence Length: 6

• CAS: 931398-72-0
• Formula: C19H16N2O5
• Solubility: DMSO: 7 mg/ml (18.96 mM); Ethanol: < 1mg/mL
• Receptor: PHD2: 21 nM
• Biological Activity: IOX2 is a selective inhibitor of the Hypoxia Inducible Factor (HIF) Prolyl-Hydroxylases (PHD); active in cells with the IC50 value of 21 nM for PHD2/ELGN-1 and no inhibition at FIH (20uM).
• Kinase Assay: Inhibition assays are carried out in 384-well white ProxiPlates in 10 uL of reaction volume. Standard reaction mixtures consisted of the compound (in 2% DMSO final concentration), enzyme mix (0.001 uM of PHD2, 10 uM of Fe(II), 100 uM of ascorbate) and peptide mix (0.06 uM of biotinylated C-terminal oxygen dependent degradation domain (CODD) peptide, 2 uM of 20G) in 50 mM HEPES pH 7.5, 0.01% Tween-20 and 0.1% BSA buffer. Compounds (e.g., IOX2) are preincubated with the enzyme mix for 15 min before being incubated with peptide mix for 10 min at 22°C. Each reaction is quenched with 5 uL of 30 mM EDTA. The bead mix containing AlphaScreen beads is preincubated for 1h with a rabbit monoclonal antibody selective for hydroxy-HIFla (Pro564) and are added to the wells for a further 1 h at 22°C. The plates are then analyzed with an Envision plate reader. The IC50 values are calculated using nonlinear regression with normalized dose-response fit using Prism GraphPad (n = or >3)[1].
• Smiles: OC(=O)CNC(=O)c1c(O)c2ccccc2n(Cc2ccccc2)c1=O
• Cell Assay: IOX2 is prepared in DMSO and stored, and then diluted with appropriate medium before use[I]. Both VHL-defective (renal carcinoma cells with an empty vector, RC(4) and VHL-competent cells human embryonic kidney HEK293T, osteosarcoma U20S and RCC4jVHLHA (RCC4 stably transfected with C-terminal HA-tagged wt VHL) are used. Cells are treated with DMSO (control) and tested compounds (e.g., IOX2) (dissolved in DMSO except for DMOG which is dissolved in PBS and added directly to culture medium) for 4-5 h. Cell extracts are probed with antibodies to hydroxyPro564 (CODD-OH) and hydroxy-Asn803 (CAD-OH). HIF-1alpha band intensities are used to normalize hydroxylation signals [1].
• Animal Experimental: Zebrafish
• Preparation and Storage: 2 years-80°C in solvent.; 3 years -20°C powder.

Structure

Related Product Information for IOX 2, inhibitor

IOX2 is a selective inhibitor of the Hypoxia Inducible Factor (HIF) Prolyl-Hydroxylases (PHD); active in cells with the IC50 value of 21 nM for PHD2/ELGN-1 and no inhibition at FIH (20uM).