A9251

Manufactured by Merck Group

Sourced in United States

The A9251 is a laboratory equipment product manufactured by Merck Group. It serves as a core component for various scientific applications, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation. The core function of the A9251 is not available for presentation at this time.

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Lab products found in correlation

Digitonin, by Merck Group (1 mentions) A1852, by Merck Group (1 mentions) Rnase 1, by Thermo Fisher Scientific (1 mentions) To pro 3 iodide, by Thermo Fisher Scientific (1 mentions) Allopurinol, by Merck Group (1 mentions) A2786, by Merck Group (1 mentions) H9377, by Merck Group (1 mentions) T7039, by Merck Group (1 mentions) M3763, by Merck Group (1 mentions) S4317, by Merck Group (1 mentions) Hy 19533, by MedChemExpress (1 mentions) Hy 100937, by MedChemExpress (1 mentions)

7 protocols using a9251

Preparation of Adenosine and 8-SPT Solutions

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The stock solution of adenosine 5′-triphosphate disodium salt hydrate (ADO; A9251, Sigma-Aldrich, St. Louis, MO) was made up as a 100 mM solution in deionized water. The stock solution of 8-(p-sulfophenyl)theophylline (8-SPT; A013, Sigma-Aldrich, St. Louis, MO) was made up as a 100 μM solution also in deionized water. The working solutions were adenosine (25μM) and 8-SPT (100 μM).

Nadal L., Garcia N., Hurtado E., Simó A., Tomàs M., Lanuza M.A., Santafé M, & Tomàs J. (2016). Presynaptic muscarinic acetylcholine autoreceptors (M1, M2 and M4 subtypes), adenosine receptors (A1 and A2A) and tropomyosin-related kinase B receptor (TrkB) modulate the developmental synapse elimination process at the neuromuscular junction. Molecular Brain, 9, 67.

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Adenosine Dose-Response Protocol for CMR

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Four doses of adenosine were used: the “standard” human dose^{35 (link)} (1x = 0.14 mg/kg/min), and double (2x = 0.28 mg/kg/min), triple (3x = 0.42 mg/kg/min), and quadruple of the standard dose (4x = 0.56 mg/kg/min). Adenosine powder (A9251, Sigma, St. Louis, Missouri, USA) was dissolved in saline at 3mg/ml, filter sterilized (VWR, 76010–404), and administered through an ear vein at the constant rate appropriate for each dose using an infusion pump. Each pig was assigned a dosing regimen with up to four doses of adenosine (Figure 1A), administered during both the CMR exam (Figure 1B) and the microsphere injection surgery (Figure 1C). Dose order was randomized for each pig to cancel out any potential biases due to previous doses or experimental timing. All procedures were executed under a strict time schedule to ensure each measurement occurred at the same time point post-infusion start from dose to dose and pig to pig, as per Figure 1. Once an infusion started, all procedures paused for two minutes to allow the hemodynamic effects to stabilize.

Weyers J.J., Ramanan V., Javed A., Barry J., Larsen M., Nayak K., Wright G.A, & Ghugre N.R. (2021). Myocardial Blood Flow is the Dominant Factor Influencing Cardiac Magnetic Resonance Adenosine Stress T2. NMR in biomedicine, 35(3), e4643.

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Sec Body Formation Regulation by Nucleotides

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Wild-type S2 cells (1.5×10⁶) were plated on coverslips and permeabilized with 10 µg/ml digitonin (D141, Sigma-Aldrich) in KRB for 2 h at 26°C. Subsequently, cells were fixed and Sec bodies were visualized by immunostaining of Sec16. To test the effect of ATP, AMP or adenosine on Sec body formation, the semi-intact cells were incubated in the presence of 0.5 mM ATP (A1852, Sigma-Aldrich), 0.5 mM AMP (01930, Sigma- Aldrich) or 0.5 mM adenosine (A9251, Sigma-Aldrich). Note that the digitonin was not removed. The permeabilization efficiency was determined by using the non-membrane-permeable dye TO-PRO-3 iodide (T3605, Thermo Fisher Scientific) (Fig. 6A). The import buffer used in the SIC system was 20 mM HEPES, 110 mM KAc, 2 mM MgAc, 5 mM NaAc and 0.5 mM EGTA (pH was set at either 6.0 or 7.4 with KOH). For the RNA degradation experiment in the SIC system, we incubated S2 cells on coverslips for 2 h at 26°C in Schneider's medium containing 10 µg/ml digitonin (Sigma-Aldrich) with or without 0.25 U/ul RNase 1 (EN0602, Thermo Fisher Scientific).

Zhang C., van Leeuwen W., Blotenburg M., Aguilera-Gomez A., Brussee S., Grond R., Kampinga H.H, & Rabouille C. (2021). Activation of IRE1, PERK and salt-inducible kinases leads to Sec body formation in Drosophila S2 cells. Journal of Cell Science, 134(17), jcs258685.

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Allopurinol and Nucleoside Metabolism in Flies

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Allopurinol (A8003, Sigma-Aldrich) was diluted in standard medium at 100 μg/ml and flies were placed for 5 d on this medium before metabolite extraction. Adenine, adenosine, and hypoxanthine (A2786, A9251, and H9377, Sigma-Aldrich) or N⁶-methyladenosine (m⁶A) (QB-1055, Combi-Blocks, San Diego, CA) were diluted in fly food medium at 500 μM. Parents were allowed to lay eggs on this medium in order to have exposition to the drug throughout all larval development of the progeny. Adults were collected and placed in normal medium until 5 d before the test, when they were placed again in food supplemented with adenosine or m⁶A at the same concentrations.

Petitgas C., Seugnet L., Dulac A., Matassi G., Mteyrek A., Fima R., Strehaiano M., Dagorret J., Chérif-Zahar B., Marie S., Ceballos-Picot I, & Birman S. (2024). Metabolic and neurobehavioral disturbances induced by purine recycling deficiency in Drosophila. eLife, 12, RP88510.

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Modulating CD73 and TGF-β Pathways

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The CD73 inhibitor adenosine 5′-(α,β-methylene) diphosphate (200 uM, M3763 Sigma-Aldrich), adenosine (10 uM, A9251 Sigma-Aldrich), TGF-β (5 ng/ml, T7039 Sigma Aldrich) and TGF-β inhibitor (1 uM, S4317 Sigma Aldrich) were added freshly to the culture media daily over 3 weeks.

Hau K.L., Ranzoni A.M., Vlahova F., Hawkins K., De Coppi P., David A.L, & Guillot P.V. (2017). TGFβ-induced osteogenic potential of human amniotic fluid stem cells via CD73-generated adenosine production. Scientific Reports, 7, 6601.

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CAR T-Cell Cytotoxicity Assay

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Triplicate wells of 5000 luciferase-expressing parental and antigen-expressing cells were co-cultured with differing ratios of CAR-expressing T cells to tumor cells as previously described (10 (link)). Cytotoxicity of T cells was evaluated the following day, and culture supernatants were collected for IFNγ ELISA (10 (link), 35 (link), 36 (link)). To evaluate the resistance of RIAD-expressing T cells to immunosuppression, in vitro cytotoxicity assays were also performed in the presence of PGE₂ (Enzo Life Sciences #BML-PG007), and adenosine (Sigma #A9251).

Newick K., O’Brien S., Sun J., Kapoor V., Maceyko S., Lo A., Pure E., Moon E, & Albelda S.M. (2016). Augmentation of CAR T cell trafficking and antitumor efficacy by blocking protein kinase A (PKA) localization. Cancer immunology research, 4(6), 541-551.

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Adenosine Receptor Modulation in Visual Flicker

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0.2 mg/ml SCH58261 [30] (A 2A receptor antagonist, HY-19533, MedChem-Express) and 0.4 mg/mL DPCPX [31] (A 1 receptor antagonist, HY-100937, MedChemExpress) were dissolved in a solution including 5% DMSO and 95% corn oil. The intragastric administrations of SCH58261 (2 mg/kg) and DPCPX (4 mg/kg) were carried out for 14 days before one-hour 40 Hz visual flicker stimulation. 5 mg/ml adenosine was dissolved in a solution including 2.5% DMSO and 97.5% saline. The intraperitoneal injection of adenosine [32] (100 mg/kg) was carried out for 7 days before behavior tests. During the in vitro electrophysiology recording, 50 mM adenosine (A9251, Sigma) was dissolved in DMSO and diluted to 10 μM in artificial cerebrospinal fluid (aCSF). DPCPX (HY-100937, MedChemExpress) at 1 mM was dissolved in DMSO and diluted in 0.3 μM in aCSF. SCH58261 (HY-19533, MedChemExpress) at 5 mM was dissolved in DMSO and diluted to 0.1 μM in aCSF. The drug administration dosage was determined based on previous studies [22, 33, 34] .

Ju J., Li X., Pan Y., Du J., Yang X., Men S., Liu B., Zhang Z., Zhong H., Mai J., Wang Y, & Hou S.T. (2024). Adenosine mediates the amelioration of social novelty deficits during rhythmic light treatment of 16p11.2 deletion female mice. Molecular psychiatry.

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