|Solubility (25°C) *||In vitro||DMSO||56 mg/mL (200.5 mM)|
|In vivo||2% DMSO+30% PEG 300+5% Tween 80+ddH2O||5mg/mL|
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
|Chemical Name||1(2H)-Isoquinolinone, 5-(2-oxo-2-phenylethoxy)-|
|Description||UPF 1069 is a selective PARP2 inhibitor with IC50 of 0.3 μM. It is ~27-fold selective against PARP1.|
|In vitro||UPF 1069 is a selective PARP2 inhibitor with IC50 of 0.3 μM while inhibiting PARP1 with IC50 of 8 μM. |
|In vivo||In organotypic hippocampal slices, PARP-2 inhibition with UPF-1069 (0.01-1 mM) causes a concentration-dependent exacerbation (up to 155%) of oxygen-glucose deprivation (OGD)-induced CA1 pyramidal cell death. Higher concentrations, acting on both PARP-1 and PARP-2, have no effect on OGD injury. In mouse mixed cortical cells exposed to OGD, UPF-1069 (1-10 mM) significantly reduces post-ischaemic damage. |
|Features||The most selective PARP2 inhibitor available to date.|
|PARP-1 and PARP-2 activity assays||PARP activity is evaluated by utilizing commercially available recombinant bovine PARP-1 and mouse PARP-2. Briefly, the enzymatic reaction is carried out in 100 mL of 50 mM Tris-HCl (pH 8.0) containing 5 mM MgCl2, 2 mM dithiothreitol, 10 mg sonicatedcalf thymus DNA, 0.2 mCi [adenine-2,8-3H]NAD and recombinant enzyme PARP-1 or PARP-2 (0.03 U per sample). Different concentrations of the putative inhibitors are added, and the mixture is incubated for 1 h at 37℃. The reaction is terminated by adding 1 mL of 10% trichloroacetic acid (w/v) and centrifuged. Pellets are then washed twice with 1 mL of H2O and resuspended in 1 mL of 0.1 M NaOH. The radioactivity incorporated from [adenine-2,8-3H]NAD into proteins is evaluated by liquid scintillation spectrometry. Nuclear extracts for PARP activity assay are prepared from PARP-1-/- and PARP-1+/+ mouse fibroblasts as previouslydescribed. Briefly, immortalized fibroblasts from PARP-1-/- and PARP-1+/+ mice are grown in DMEM supplemented with 2 mM glutamine, 10% bovine serum and antibiotics at 37℃. Nuclear extracts are prepared as follows: 10 cm confluent plates are scraped in 1 mL of homogenization buffer composed of 75 mM sucrose, 225 mM mannitol and 5 mM Tris, pH 7.4 plus 10 mL protease inhibitor cocktail. The cell suspension is homogenized with a Teflon pestle, and the mixture is centrifuged at 600 × g for 5 min at 4℃. The crude nuclear pellet is washed and resuspended in 1 mL of PARP assay buffer (5 mM MgCl2, 2 mM DTT, 50 mM Tris, pH 8) containing 100 mM N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) to fully activate PARP activity. Samples containing 100 mL of the resuspended nuclear pellet are incubated for 60 min at 37℃ in the presence of 35.5 nM 3H-NAD. The reaction is stopped with 1 mL of 10% trichloroacetic acid (w/v), and the mixture is centrifuged at 12 000 × g for 10 min at 4℃. The reaction is terminated by the addition of 1 mL of 10% trichloroacetic acid (w/v), and radioactivity of the suspension is measured by liquid scintillation spectrometry.|
|Animal Models||Rats deprived of hippocampi.|
PLEASE KEEP THE PRODUCT UNDER -20°C FOR LONG-TERM STORAGE.
NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.
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