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4-Thio-UTP

Catalog No.GB20114

4-Thio-UTP is a UTP analog and potent P2Y2 and P2Y4 agonist with EC50 values of 35 and 350nM for hP2Y2 and hP2Y4, respectively.

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4-Thio-UTP Chemical Structure

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10ul (100mM)
$155.00
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50ul (100mM)
$742.00
In stock
100ul (100mM)
$1,361.00
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Sample solution is provided at 25 µL, 10mM.

Description of 4-Thio-UTP

4-Thio-UTP is a UTP analog and potent P2Y2 and P2Y4 agonist with EC50 values of 35 and 350nM for hP2Y2 and hP2Y4, respectively[1]. P2Y2 and P2Y4 are G-protein-coupled purinergic receptors activated extracellularly by UTP that mobilize intracellular Ca²⁺ and thereby regulate epithelial ion/water transport, immune cell recruitment and vascular tone[2]. 4-Thio-UTP is widely used to investigate purinergic signaling in inflammation, epithelial ion transport, neurotransmission, and immune regulation[3]. 4-Thio-UTP is also used in cross-linking experiments and for labeling transcription complexes to explore the behavior of RNA polymerase during transcription elongation and termination[4].

In vitro, 4-Thio-UTP (0.1-300μM; 20min) evoked a concentration-dependent accumulation of [³H]inositol polyphosphates in cultured rat aortic smooth muscle cells[5].

In vivo, 4-Thio-UTP (100μM; 15min; basolateral perfusion) stimulated chloride secretion in wild-type mouse jejunum and the response was attenuated in both P2Y4-and P2Y2-deficient mice[6]. 4-Thio-UTP (0.44mg/kg; i.v.; 30min before LAD ligation) reduced myocardial infarct size and improved left-ventricular fractional shortening in wild-type mice but lost protection in P2Y2⁻/⁻mice[7].

References:
[1] Jacobson KA, Costanzi S, Ivanov AA, et al. Structure activity and molecular modeling analyses of ribose- and base-modified uridine 5'-triphosphate analogues at the human P2Y2 and P2Y4 receptors. Biochem Pharmacol. 2006;71(4):540-549.
[2] Hede SE, Amstrup J, Klaerke DA, Novak I. P2Y2 and P2Y4 receptors regulate pancreatic Ca(2+)-activated K+ channels differently. Pflugers Arch. 2005;450(6):429-436.
[3] Lazarowski ER, Boucher RC. UTP as an extracellular signaling molecule. News Physiol Sci. 2001;16:1-5.
[4] Bartholomew B, Dahmus ME, Meares CF. RNA contacts subunits IIo and IIc in HeLa RNA polymerase II transcription complexes. J Biol Chem. 1986;261(30):14226-14231.
[5] Kumari R, Goh G, Ng LL, Boarder MR. ATP and UTP responses of cultured rat aortic smooth muscle cells revisited: dominance of P2Y2 receptors. Br J Pharmacol. 2003;140(7):1169-1176.
[6] Ghanem E, Robaye B, Leal T, et al. The role of epithelial P2Y2 and P2Y4 receptors in the regulation of intestinal chloride secretion. Br J Pharmacol. 2005;146(3):364-369.
[7] Cohen R, Shainberg A, Hochhauser E, et al. UTP reduces infarct size and improves mice heart function after myocardial infarct via P2Y2 receptor. Biochem Pharmacol. 2011;82(9):1126-1133.

Protocol of 4-Thio-UTP

Cell experiment [1]:

Cell lines

rat aortic smooth muscle cells

Preparation Method

Rat aortic VSMCs from both spontaneously hypertensive (SHR) and normotensive (WKY) animals were prepared according to protocol. For the total [3H]-inositol (poly)phosphate measurements, cells were culturedto 80% confluence in 24-well multi-well plates. The medium was then replaced with 500μl of serumfree M199 medium supplemented with 50IU/ml penicillin, 50μg/ml streptomycin, 2mM L-glutamine and1μCi/ml(0.037MBq/ml) D-myo-[2-3H]inositol. After 24h, cells were incubated with 10mM LiCl for 10min – where appropriate suramin was included with the lithium. After 20min stimulation with 4-Thio-UTP (0.1-300μM) in the continuing presence of lithium, the reaction was stoppedwith trichloroacetic acid, the solution was neutralised with freon : octylamine extraction, and addition of NaHCO3 and[3H]inositol phosphates separatedon small Dowex-1 columns.

Reaction Conditions

0.1-300μM; 20min

Applications

4-Thio-UTP evoked a concentration-dependent accumulation of [³H]inositol polyphosphates in cultured rat aortic smooth muscle cells.

Animal experiment [2]:

Animal models

P2Y2⁻/⁻ and P2Y4⁻/⁻ mice; DF508 mice

Preparation Method

P2Y2⁻/⁻ and P2Y4⁻/⁻ mice and their littermates were maintained on a standard diet in conventional facilities. DF508 mice and their littermates were weaned to a liquid diet and the colony was maintained in a pathogen-free status. Mice were kept in a 12-h light–dark cycle. chloride secretion was assessed as the Isc following addition of phloridzin or amiloride to the apical side as well as of indomethacin (100μM) to both bathing media. 4-Thio-UTP was added, at 100μM, to the apical or basolateral solution, unless otherwise stated. Forskolin was always added to the basolateral solution at the concentration of 10μM in ethanol 0.1%; this concentration of ethanol does not affect Isc. The increase in Isc was calculated as the difference between the basal current and the peak current obtained within 15min of addition of 4-Thio-UTP or forskolin. Statistical analysis was performed using the unpaired t-test and a P-value<0.05 was considered significant.

Dosage form

100μM; 15min; basolateral perfusion

Applications

4-Thio-UTP stimulated chloride secretion in wild-type mouse jejunum and the response was attenuated in both P2Y4- and P2Y2-deficient mice.

References:
[1] Kumari R, Goh G, Ng LL, Boarder MR. ATP and UTP responses of cultured rat aortic smooth muscle cells revisited: dominance of P2Y2 receptors. Br J Pharmacol. 2003;140(7):1169-1176.
[2] Ghanem E, Robaye B, Leal T, et al. The role of epithelial P2Y2 and P2Y4 receptors in the regulation of intestinal chloride secretion. Br J Pharmacol. 2005;146(3):364-369.

Chemical Properties of 4-Thio-UTP

Cas No. SDF
Formula C9H15N2O14P3S (free acid) M.Wt 500.2 (free acid)
Solubility Storage Store at -20°C or below
General tips Please select the appropriate solvent to prepare the stock solution according to the solubility of the product in different solvents; once the solution is prepared, please store it in separate packages to avoid product failure caused by repeated freezing and thawing.Storage method and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored at -20°C, please use it within 1 month.
To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time.
Shipping Condition Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request.

Complete Stock Solution Preparation Table of 4-Thio-UTP

Prepare stock solution
1 mg 5 mg 10 mg
1 mM 1.9992 mL 9.996 mL 19.992 mL
5 mM 399.8 μL 1.9992 mL 3.9984 mL
10 mM 199.9 μL 999.6 μL 1.9992 mL
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