The aCSF used for dissections and recordings contained (in mM) 127 NaCl, 26 NaHCO3, 10 glucose, 3 KCl (unless otherwise stated), 2 CaCl, 1.25 NaH2PO4, and 1 MgCl2. TFLLR, theophylline, MSO, FA and glutamine were supplied by Sigma-Aldrich (Poole, UK); DPCPX and SCH58261 were supplied by Abcam (Cambridge, UK); ARL67156 was supplied by Tocris Bioscience (Bristol, UK). All drugs were dissolved in reverse-osmosis water, except picrotoxin, DPCPX and SCH58261, which were dissolved in DMSO. The concentration of DMSO in working solutions did not exceed 0.1% (v/v).
Sch58261
Manufactured by Abcam
Sourced in United Kingdom
SCH58261 is a potent and selective adenosine A2A receptor antagonist. It exhibits high affinity and specificity for the adenosine A2A receptor.
Automatically generated - may contain errors
Lab products found in correlation
Theophylline, by Merck Group (1 mentions)
Glutamine, by Merck Group (1 mentions)
Tfllr, by Merck Group (1 mentions)
Dpcpx, by Abcam (1 mentions)
Arl67156, by Bio-Techne (1 mentions)
7 aad, by Miltenyi Biotec (1 mentions)
Psb603, by Bio-Techne (1 mentions)
Bay 60 6583, by Bio-Techne (1 mentions)
Mre 3008f20, by Bio-Techne (1 mentions)
5 n ethylcarboxamidoadenosine neca, by Bio-Techne (1 mentions)
Cgs21680, by Abcam (1 mentions)
4 protocols using sch58261
1
Extracellular Solutions Composition
2
Assessing Spontaneous Locomotor Activity
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To evaluate the spontaneous locomotor activity, we performed the open field test. In brief, the mice were placed in the center of an activity field arena (30 × 30 cm, surrounded by four 50 cm high black painted walls) equipped with a camera above to record activity. Mice were administered (ip) with vehicle or SCH58261 (3.75 mg/kg; Abcam Biochemicals, Cambridge, UK) 15 min before measuring the exploratory behavior of the animals during a 10-min period. The total distance travelled and the activity within the outer and inner zone of the open field was analyzed using Spot tracker function from Image J (NIH). All behavioral tests were carried out in a sound attenuated room with 15 lux illumination. The apparatus and the objects were cleaned with a 70% alcohol solution and rinsed with water after each session.
3
Cytotoxic Assay of CAR T Cells
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HeLa cells were labeled with PKH26 dye, to distinguish the target cells from T cells. 3 × 104 labeled HeLa cells were co-incubated at 1:1, 1:5, 1:10, and 1:20 ratios with CAR T, Un-T, and Mock T cells for 18 h. Also, 1 μM of the A2aR antagonist SCH58261(Abcam, Cambridge, MA) in the presence and absence of 1 μM 5′-(N-ethylcarboxamido) adenosine (NECA- Adenosine receptor agonist) (Abcam). After 18 h of co-incubation, cells were stained with 7-AAD (Miltenyi Biotec) as a viability dye to exclude dead cells and analyzed by flow cytometry. PKH26 positive /7AAD positive population demonstrated dead tumor cells. To calculate the percentage of dead tumor cells, the percentage of spontaneous lysis of target cells which were incubated in the absence of effector cells, were subtracted from the percentage of target cells that were incubated with effector cells. FlowJo (v7.6.1) software was used for data analysis.
4
Adenosine Receptor Ligands and Analogs
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The adenosine A1 receptor agonist N6-cyclopentyladenosine (N6-CPA, purity > 98%) was obtained from Abcam (Shanghai, China) and dissolved in DMSO at 50 mM. The adenosine A1 receptor antagonist rolofylline (KW-3902, purity > 99%) was obtained from Tocris Bioscience (Bristol, United Kingdom) and dissolved in DMSO at 10 mM. The adenosine A2a receptor agonist CGS 21680 (purity > 98%) was obtained from Abcam and dissolved in DMSO at 10 mM. The adenosine A2a receptor antagonist SCH 58261 (purity > 99%) was obtained from Abcam and dissolved in DMSO at 10 mM. The adenosine A2b receptor agonist BAY 60-6583 (purity > 98%) was obtained from Tocris Bioscience and dissolved in DMSO at 20 mM. The adenosine A2b receptor antagonist PSB603 (purity > 98%) was obtained from Tocris Bioscience and dissolved in DMSO at 10 mM. The adenosine A3 receptor agonist piclidenoson (IB-MECA, purity > 97%) was obtained from Abcam and dissolved in DMSO at 10 mM. The adenosine A3 receptor antagonist MRE 3008F20 (purity > 98%) was obtained from Tocris Bioscience and dissolved in DMSO at 50 mM. The adenosine analog 5′-N-ethylcarboxamidoadenosine (NECA, purity > 99%) was obtained from Tocris Bioscience and dissolved in DMSO at 50 mM. All solutions were stored at –20°C and used within 1 year.
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