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Ehna hydrochloride

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EHNA hydrochloride is a chemical compound used in laboratory research. It functions as an inhibitor of the enzyme adenosine deaminase. The core purpose of EHNA hydrochloride is to facilitate scientific investigations and experiments in controlled laboratory settings.

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Lab products found in correlation

Adenosine assay kit, by Abcam (4 mentions) Tr717, by Thermo Fisher Scientific (2 mentions) Enliten atp assay system, by Promega (2 mentions) Victor nivo, by PerkinElmer (1 mentions) Infinite 200 pro, by Tecan (1 mentions) Isoproterenol hydrochloride, by Merck Group (1 mentions) Vinpocetine, by Merck Group (1 mentions) 3 isobutyl 1 methyl xanthine (ibmx), by Santa Cruz Biotechnology (1 mentions) Penicillin streptomycin, by Merck Group (1 mentions) M199 medium, by Thermo Fisher Scientific (1 mentions) Mouse α actinin, by Merck Group (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Laminin, by Merck Group (1 mentions) Creatine monohydrate, by Merck Group (1 mentions) Cilostamide, by Bio-Techne (1 mentions) Carnitine hydrochloride, by Merck Group (1 mentions) Tadalafil, by Santa Cruz Biotechnology (1 mentions) Dipyridamole, by Merck Group (1 mentions) Pertussis toxin, by Bio-Techne (1 mentions) Mrs1220, by Bio-Techne (1 mentions)

Spelling variants of this product

Hydrochlorides (2 citations) Erythro-9-(2-Hydroxy-3-nonyl)-adenine hydrochloride (EHNA, (2 citations)

7 protocols using ehna hydrochloride

1

Hippocampal Adenosine Quantification

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One day after the behavioral tests, hippocampal samples were collected and subjected to test the adenosine level. Hippocampal samples were lysed with RIPA with protein inhibitors and adenosine deminase inhibitor EHNA hydrochloride (E114, Sigma, St. Louis, MO, USA). Then, hippocampal adenosine is measured using the Adenosine Assay Kit (K327-100, BioVision, Milpitas, CA, USA). Fluorescence was measured using a multimode plate reader (VICTOR Nivo, PerkinElmer, Waltham, MA, USA). Adenosine in samples was calculated based on a calibration curve from standard adenosine samples, and the hippocampal adenosine level was expressed as 100% of the control group.
Liu Y., Du T., Zhang W., Lu W., Peng Z., Huang S., Sun X., Zhu X., Chen C., Qian L., Wen L., Xu P, & Zhang Y. (2019). Modified Huang-Lian-Jie-Du Decoction Ameliorates Aβ Synaptotoxicity in a Murine Model of Alzheimer's Disease. Oxidative Medicine and Cellular Longevity, 2019, 8340192.
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2

Quantifying ATP and Adenosine Levels

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ATP is measured by a luciferase reaction where light (560 nm) was emitted when D-luciferin is converted to oxyluciferin using the ENLITEN ATP Assay System (FF2000, Promega)8 , 12 . To inhibit ATP hydrolysis, condition media or microdialysis samples were incubated with the ectonucleotidase inhibitor ARL 67156 (6-N, N-diethyl- β-γ-dibromo-methylene-d- adenosine- 5-triphosphate trisodium salt hydrate, FPL 67156). Luminescence was measured using a luminometer (PE Applied Biosystems, TR717). ATP in samples was calculated based on a calibration curve with standard ATP samples.
Adenosine is measured by using the Adenosine Assay Kit (K327-100, BioVision). To inhibit adenosine degradation, samples were incubated with adenosine deminase inhibitor EHNA hydrochloride (E114, Sigma). Fluorescence was measured using a luminometer (PE Applied Biosystems, TR717). Adenosine in samples was calculated based on a calibration curve from standard adenosine samples.
Sun X.D., Li L., Liu F., Huang Z.H., Bean J.C., Jiao H.F., Barik A., Kim S.M., Wu H., Shen C., Tian Y., Lin T.W., Bates R., Sathyamurthy A., Chen Y.J., Yin D.M., Xiong L., Lin H.P., Hu J.X., Li B.M., Gao T.M., Xiong W.C, & Mei L. (2016). Lrp4 in astrocytes modulates glutamatergic transmission. Nature neuroscience, 19(8), 1010-1018.
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3

Quantifying ATP and Adenosine Levels

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ATP is measured by a luciferase reaction where light (560 nm) was emitted when D-luciferin is converted to oxyluciferin using the ENLITEN ATP Assay System (FF2000, Promega)8 , 12 . To inhibit ATP hydrolysis, condition media or microdialysis samples were incubated with the ectonucleotidase inhibitor ARL 67156 (6-N, N-diethyl- β-γ-dibromo-methylene-d- adenosine- 5-triphosphate trisodium salt hydrate, FPL 67156). Luminescence was measured using a luminometer (PE Applied Biosystems, TR717). ATP in samples was calculated based on a calibration curve with standard ATP samples.
Adenosine is measured by using the Adenosine Assay Kit (K327-100, BioVision). To inhibit adenosine degradation, samples were incubated with adenosine deminase inhibitor EHNA hydrochloride (E114, Sigma). Fluorescence was measured using a luminometer (PE Applied Biosystems, TR717). Adenosine in samples was calculated based on a calibration curve from standard adenosine samples.
Sun X.D., Li L., Liu F., Huang Z.H., Bean J.C., Jiao H.F., Barik A., Kim S.M., Wu H., Shen C., Tian Y., Lin T.W., Bates R., Sathyamurthy A., Chen Y.J., Yin D.M., Xiong L., Lin H.P., Hu J.X., Li B.M., Gao T.M., Xiong W.C, & Mei L. (2016). Lrp4 in astrocytes modulates glutamatergic transmission. Nature neuroscience, 19(8), 1010-1018.
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4

Cardiac Tissue Metabolite Profiling Post-MI/R

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Rat hearts were harvested at 1 and 3 days post MI/R injury. Hearts were rapidly perfused with adenosine deaminase inhibitor, EHNA hydrochloride (5 μmol/L, Sigma), and ligated from its attachment to the great vessels. Atria were excised just below the level of the appendages and the right ventricular free wall was removed. LV tissue was then snap frozen in liquid nitrogen, and kept at −80°C until further processing. Preparation of myocardial tissue for LC/MS was performed as previously described with the following modifications.36, 37 Frozen tissue was homogenized in prechilled acetonitrile/methanol/water (1:2:2, vol/vol/vol). Crude lysate was then heated for 10 minutes at 60°C, cooled on ice for 10 minutes, and then centrifuged at 2350g for 90 minutes at 4°C. Supernatant was isolated and further purified by centrifugation at 21 000g for 3 minutes through a 35‐μm filter. Supernatant was then diluted 1:1000 in double‐distilled H2O to prevent mass overload and then taken to LC/MS for metabolite analysis.
Shin E.Y., Wang L., Zemskova M., Deppen J., Xu K., Strobel F., García A.J., Tirouvanziam R, & Levit R.D. (2018). Adenosine Production by Biomaterial‐Supported Mesenchymal Stromal Cells Reduces the Innate Inflammatory Response in Myocardial Ischemia/Reperfusion Injury. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 7(2), e006949.
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5

Quantifying Adenosine Levels with Assay

Cited 1 time
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Adenosine test was performed as described previously [25 (link)]. In brief, adenosine is measured with the Adenosine Assay Kit (K327-100, BioVision). Samples were incubated with adenosine deminase inhibitor EHNA hydrochloride (E114, Sigma) to inhibit adenosine degradation. Fluorescence was measured using a microplate reader (TECAN, Infinite 200 PRO). Adenosine in samples was calculated based on a calibration curve from standard adenosine samples.
Liu Z.Y., Li Y.Q., Wang D.L., Wang Y., Qiu W.T., Qiu Y.Y., Zhang H.L., You Q.L., Liu S.M., Liang Q.N., Wu E.J., Hu B.J, & Sun X.D. (2024). Agrin-Lrp4 pathway in hippocampal astrocytes restrains development of temporal lobe epilepsy through adenosine signaling. Cell & Bioscience, 14, 66.
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6

Cardiomyocyte contractility regulation

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M199 medium (Invitrogen UK, 11150), taurine (Biochemica, A1141), creatine monohydrate (Sigma Aldrich, C3630), penicillin/streptomycin (Merck, A2212), carnitine hydrochloride (Sigma Aldrich, C9500) BSA (Sigma Aldrich, A6003), laminin (Sigma Aldrich L2020), isoproterenol hydrochloride (Sigma Aldrich, I6504), ICI118551 (Tocris UK, 0821), CGP 20712A (Tocris UK, 1024), SR 59230A hydrochloride (Tocris UK, 1511), L-NAME hydrochloride (Tocris UK, 0665), Vinpocetine (Sigma Aldrich, V6383), EHNA hydrochloride (Sigma Aldrich, E114), Cilostamide (Tocris UK, 0915), Tadalafil (Santa Cruz USA, sc-208412), IBMX (Santa Cruz sc-201188), self-made rabbit sGCα and β subunit antibodies (specificity tested in KO animals; Friebe et al., 2018 (link)), mouse α-actinin (Sigma Aldrich, A7732), mouse Caveolin-3 (BD Transduction Laboratories, 610421, specificity tested in KO animals; Woodman et al., 2002 (link)), secondary Alexa Fluor antibodies 488 nm, 514 nm, 568 nm and 633 nm (Life Technologies), BSA (Fisher Scientific UK, BPE9704), fluorescence mounting medium (Vectashield Germany, H-1000), MaTek glass-bottom dishes (MaTek USA, P35G-1.5–10 C), TAT-scram and TAT-Cav3 peptides (a gift from Dr. Sarah Calaghan from Leeds, England).
Schobesberger S., Wright P.T., Poulet C., Sanchez Alonso Mardones J.L., Mansfield C., Friebe A., Harding S.E., Balligand J.L., Nikolaev V.O, & Gorelik J. (2020). β3-Adrenoceptor redistribution impairs NO/cGMP/PDE2 signalling in failing cardiomyocytes. eLife, 9, e52221.
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7

Adenosine Receptor Signaling Assay

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Adenosine, AMP (Adenosine 5’-monophosphate disodium salt), DPCPX (8-cyclopentyl-1,3-dipropylxanthine), EHNA hydrochloride, pentostatin (dCF), and dipyridamole (DPM) were obtained from Sigma Aldrich (St Louis, MO). PD 184352 (2-[(2-chloro-4-iodophenyl)amino]-N-cyclopropylmethoxy)-3,4-difluorobenzamide), U 73122 (1-[6-[[(17β)-3-methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-1H-pyrrole -2,5-dione), ANR 94 (8-ethoxy-9-ethyl-9H-purin-6-amine), PSB 603 (8-[4-[4-(4-Chloro phenzyl)piperazide-1-sulfonyl)phenyl]]-1-propylxanthine), MRS 1220 (N-[9-chloro-2-(2-fura nyl)[1 (link),2 (link),4 ]-triazolo[1,5-c]quinazolin-5-yl]benzene acetamide), pertussis toxin (50 ng/ml), IB-MECA (1-deoxy-1-[6-[[(3-iodophenyl)methyl]amino]-9H-purin-9-yl]-N-methyl-β-D-ribofuranuronamide), and CPA (N-cyclopentylAdenosine) were purchased from Tocris (Ellisville, MO). SDF-1 was from PeproTech (London, UK).
Tarnowski M., Tkacz M., Piotrowska K., Zgutka K, & Pawlik A. (2018). Differential effect of adenosine on rhabdomyosarcoma migration and proliferation. Archives of Medical Science : AMS, 16(2), 414-427.
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