Neuro-2 A neuroblastoma (N2A) cells stably expressing YFP-tagged human tau cDNA (N2A-tau) driven by the CMV promoter were prepared in house, as previously described (16 (link)). Cells were cultured in Dulbecco’s modified Eagle medium supplemented with 10% fetal bovine serum, 100 U/mL streptomycin (Cellgro, Herdon, VA) and 100 mg/mL Hygromycin B (Invitrogen, Carlsbad, CA) at 37°C in the presence of 5% CO2 as previously described (17 (link)). The cells were cultured to 80% to 90% confluence in six-well plates and then changed to fresh medium containing 50 μM DL-homocysteine (Sigma, St Louis, MO), 40μM adenosine (Sigma, St. Louis, MO), and 10μM erythro-9-(2-hydroxy-3-nonyl)-adenine hydrochloride (Sigma, St. Louis, MO), or 100 μM zileuton (Sigma, St Louis, MO), as previously described (11 (link), 13 (link)). After 24 hrs, cell lysates harvested and protein extracts used for Western blotting analyses.
Erythro 9 2 hydroxy 3 nonyl adenine hydrochloride
Manufactured by Merck Group
Sourced in Australia
Erythro-9-(2-hydroxy-3-nonyl)-adenine hydrochloride is a laboratory chemical compound. It is a derivative of adenine with a nonyl group attached at the 9-position and a hydroxy group at the 2-position. The compound is available for use in research and analytical applications.
Automatically generated - may contain errors
Lab products found in correlation
Adenosine, by Merck Group (3 mentions)
Streptomycin, by Corning (3 mentions)
Hygromycin b, by Thermo Fisher Scientific (2 mentions)
Zileuton, by Merck Group (2 mentions)
Dl homocysteine, by Merck Group (2 mentions)
Lithium heparin tube, by BD (2 mentions)
Lithium heparin tubes, by Merck Group (2 mentions)
Dipyridamole, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Lipopolysaccharide, by Merck Group (1 mentions)
Colorimetric assay kit, by Cell Biolabs (1 mentions)
Dl hcy, by Merck Group (1 mentions)
6 protocols using erythro 9 2 hydroxy 3 nonyl adenine hydrochloride
1
Neuroblastoma Cell Line for Tau Aggregation
2
Infant Blood Processing for cAMP Measurement
3
Lipopolysaccharide-Induced Inflammation Assay
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Lipopolysaccharide (from Escherichia coli, serotype 055:B5), AMP, adenosine 5′‐(α, β‐methylene) diphosphate (APCP), erythro‐9‐(2‐hydroxy‐3‐nonyl)−adenine hydrochloride, and dimethyl sulfoxide were purchased from Sigma (St. Louis, MO). ZM 241385 was obtained from Tocris Bioscience/Bio‐Techne. When used as a solvent, final dimethyl sulfoxide concentrations in all assays did not exceed 0.1%; and the same dimethyl sulfoxide concentrations were used in vehicle controls.
4
Neuroblastoma Model for Alzheimer's Research
5
Neuroblastoma Cell Line for Tau Aggregation
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Neuro-2 A neuroblastoma (N2A) cells stably expressing YFP-tagged human tau cDNA (N2A-tau) driven by the CMV promoter were prepared in house, as previously described (16 (link)). Cells were cultured in Dulbecco’s modified Eagle medium supplemented with 10% fetal bovine serum, 100 U/mL streptomycin (Cellgro, Herdon, VA) and 100 mg/mL Hygromycin B (Invitrogen, Carlsbad, CA) at 37°C in the presence of 5% CO2 as previously described (17 (link)). The cells were cultured to 80% to 90% confluence in six-well plates and then changed to fresh medium containing 50 μM DL-homocysteine (Sigma, St Louis, MO), 40μM adenosine (Sigma, St. Louis, MO), and 10μM erythro-9-(2-hydroxy-3-nonyl)-adenine hydrochloride (Sigma, St. Louis, MO), or 100 μM zileuton (Sigma, St Louis, MO), as previously described (11 (link), 13 (link)). After 24 hrs, cell lysates harvested and protein extracts used for Western blotting analyses.
6
Infant Blood Processing for cAMP Measurement
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Infant cord and peripheral blood (800 l) for whole blood stimulation and white blood cell extended differential counts was collected by venipuncture or from an existing umbilical catheter (DOL 1 only) on days 7, 14, 21 and 28 into lithium-heparin tubes (Becton Dickinson, North Ryde, Australia).
An additional 400 l of infant blood was collected, as above, into lithium-heparin tubes containing 2 µl of 2mM dipyridamole (in DMSO; Sigma-Aldrich, Castle Hill, Australia) and 2 µl of 200 µM erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride (in water; Sigma-Aldrich). Samples were centrifuged at 6,000 x g for 2 minutes and plasma was removed. Blood cell pellets were resuspended in 1 ml 0.1M HCL and incubated at room temperature for 20 minutes. These samples were centrifuged at 1,000 x g for 10 minutes and supernatant (blood lysate) was stored at −80°C until cAMP measurement.
An additional 400 l of infant blood was collected, as above, into lithium-heparin tubes containing 2 µl of 2mM dipyridamole (in DMSO; Sigma-Aldrich, Castle Hill, Australia) and 2 µl of 200 µM erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride (in water; Sigma-Aldrich). Samples were centrifuged at 6,000 x g for 2 minutes and plasma was removed. Blood cell pellets were resuspended in 1 ml 0.1M HCL and incubated at room temperature for 20 minutes. These samples were centrifuged at 1,000 x g for 10 minutes and supernatant (blood lysate) was stored at −80°C until cAMP measurement.
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