Adenosine ado

Manufactured by Merck Group

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Adenosine (Ado) is a nucleoside compound that plays a crucial role in various cellular processes. It is a naturally occurring substance found in all living cells and is involved in energy metabolism, signal transduction, and cellular regulation. Adenosine serves as a building block for the synthesis of adenosine triphosphate (ATP), the primary energy currency of cells. Additionally, it acts as a neuromodulator, influencing neurotransmission and various physiological functions.

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Lab products found in correlation

4 protocols using adenosine ado

CD73 Activity Assay with HPLC

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In this assay, all reagents including tiamulin hydrogen fumarate (THF), Adenosine monophosphate (AMP) and Adenosine (Ado) were purchased from Sigma-Aldrich (St Louis, MO, USA), except Recombinant Human 5′-Nucleotidase/CD73 Protein (rhCD73) (R&D Systems, MN, USA). CD73 activity was analyzed by measuring the conversation of AMP to Ado with high performance liquid chromatography (HPLC) system (Agilent Technologies Inc., CA, USA). Briefly, rhCD73 (100 ng/mL) was prepared in assay buffer (25 mM Tris, 5 mM MgCl2, pH 7.5). Then, assay buffer alone (control) or with THF (5, 10 or 20 μM) was added. After 10 min, AMP (100 μM) was added for an additional 10 min. The production of Ado was detected by HPLC at 254 nm, and CD73 activity was expressed as Ado production per mg protein in 10 min. As well as THF, α, β-Methylene adenosine-5′-disphosphate (APCP, a specific inhibitor of CD73) (10 μM) or tylosin (an antibiotic similar to THF in antibiotic spectrum) (10 μM) was used in this assay as positive or negative control.

Yang X., Pei S., Wang H., Jin Y., Yu F., Zhou B., Zhang H., Zhang D, & Lin D. (2017). Tiamulin inhibits breast cancer growth and pulmonary metastasis by decreasing the activity of CD73. BMC Cancer, 17, 255.

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Intrathecal and Intraplantar Adenosine Delivery

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Adenosine (Ado), AMP and CPA (direct A₁R agonist) were purchased from Sigma; hPAP was graciously provided by Dr. Mark Zylka (University of North Carolina). 5’-iodotubercidin (ITU; adenosine kinase inhibitor) was purchased from Enzo Life Sciences (Farmingdale, NY). Adenosine (Ado), AMP and CPA were dissolved in 0.9% saline at pH 7.4. AMP was delivered at a concentration of 200 nmol/10 µl (Sowa et al., 2010b (link)); Ado and CPA were delivered at a dose of 10 nmol/10 µl (DeLander and Hopkins, 1987 (link), Lima et al., 2010 (link)); hPAP was delivered at a dose of 250 mU/10 µL (Zylka et al., 2008 (link)). ITU was co-administered with Ado or AMP at a concentration of 5 nmol/10 µL (Sowa et al., 2010c (link)). Intrathecal injections (hPAP, AMP + ITU, Ado + ITU, CPA) were performed using a 28 gauge, 8 mm insulin syringe (BD Biosciences, San Jose, CA) using the direct lumbar puncture method between L5-L6 (Fairbanks, 2003 (link)). Intraplantar (i.pl) injections (AMP + ITU, Ado + ITU, CPA) were performed using a 31 gauge, ½” insulin syringe (BD Biosciences) between the distal volar footpads of the right hind paw (Hurt and Zylka, 2012 (link)).

Katz N.K., Ryals J.M, & Wright D.E. (2014). Central or Peripheral Delivery of an Adenosine A1 Receptor Agonist Improves Mechanical Allodynia in a Mouse Model of Painful Diabetic Neuropathy. Neuroscience, 285, 312-323.

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Signaling Cascades in Neuronal Differentiation

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High-glucose Dulbecco’s modified Eagle’s medium (DMEM) and Opti-MEM/GlutaMAX were from Gibco (Thermo Scientific, Waltham, MA, USA)); phosphate-buffered saline (pH 7.4), L-glutamine, antibiotics, (penicillin/streptomycin mixture, G-418), donor horse serum, and fetal bovine serum (FBS) were from Biowest (Nuaille, France). 3-(4,5-dimethylthiazol-2-yl)-2–5-diphenyltetrazolium bromide (MTT) and adenosine (Ado) were purchased from Sigma (St. Louis, MO, USA). Reagents for SDS-PAGE were from Bio-Rad (California, USA). NGF (from mouse submaxillary glands) and BDNF Emax ImmunoAssay System were from Promega (Madison, USA). PageRuler™ Plus Prestained Protein Ladder (10–250 kDa) was obtained from Thermo Scientific (Waltham, MA, USA). Anti ERK 1/2, anti-phospho ERK 1/2, anti-CREB, anti-phospho-CREB, anti-Akt, anti-phospho-Akt, anti-NF-қB, anti-phospho-NF-қB, anti-β-actin monoclonal antibodies, and anti-rabbit IgG AP-linked antibody were obtained from Cell Signaling Technology (Leiden, The Netherlands). Anti-CPE monoclonal antibody was from Invitrogen (Massachusetts, USA). Anti-BDNF polyclonal antibody was from ABclonal (MA, USA). 5-Bromo4-chloro-3-indolyl phosphate disodium salt (BCIP) and nitro blue tetrazolium (NBT) were from Carl Roth GmbH (Karlsruhe, Germany). LY294002 was obtained from MedChemExpress (New York, USA).

Kazana W., Jakubczyk D., Siednienko J., Zambrowicz A., Macała J, & Zabłocka A. (2023). Mechanism of Molecular Activity of Yolkin—a Polypeptide Complex Derived from Hen Egg Yolk—in PC12 Cells and Immortalized Hippocampal Precursor Cells H19-7. Molecular Neurobiology, 60(5), 2819-2831.

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Multiplexed Neurochemical Quantification

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Fluorescein, fluorinert FC-40, dopamine hydrochloride,
acetylcholine chloride, dl-norepinephrine hydrochloride,
serotonin hydrochloride, adenosine (Ado), and γ-aminobutyric
acid (GABA) were purchased from Sigma-Aldrich (St. Louis, MO). Solutions
were prepared with HPLC-grade water. For the multiplexing detection
experiment, a master solution of neurochemical standards was prepared
by dissolving six neurochemicals into deionized (DI) water, including
25 μM Ado, 50 μM acetylcholine (ACh), 50 μM dopamine
(DA), 125 μM norepinephrine (NE), and 125 μM serotonin
(5-HT). Then, this mixture was diluted into a series of concentrations
while GABA was kept at 25 μM in all mixtures.

Zhang Y., Li K., Zhao Y., Shi W., Iyer H., Kim S., Brenden C., Sweedler J.V, & Vlasov Y. (2022). Attomole-Level Multiplexed Detection of Neurochemicals in Picoliter Droplets by On-Chip Nanoelectrospray Ionization Coupled to Mass Spectrometry. Analytical Chemistry, 94(40), 13804-13809.

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