Human fibronectin was from Merck Millipore. Ionomycin was from Apollo Scientific (Stockport, UK). Rapamycin was from Cambridge Bioscience (Cambridge, UK). SNAP substrates were from New England Biolabs (Hitchin, UK). Other reagents, including histamine and nigericin, were from Sigma-Aldrich.
Ionomycin
Manufactured by Apollo Scientific
Sourced in United Kingdom
Ionomycin is a calcium ionophore that facilitates the movement of calcium ions across cell membranes. It is commonly used in cell biology research to study calcium signaling pathways and to stimulate the activation of various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Nigericin, by Merck Group (1 mentions)
Human fibronectin, by Merck Group (1 mentions)
Histamine, by Merck Group (1 mentions)
Rapamycin, by Cambridge Bioscience (1 mentions)
Escherichia coli bl21, by GE Healthcare (1 mentions)
Recombinant human rh il 4, by R&D Systems (1 mentions)
Rh granulocyte macrophage colony stimulating factor rhgm csf, by R&D Systems (1 mentions)
Rhtnf α, by R&D Systems (1 mentions)
Pgex 6p 1, by GE Healthcare (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Phorbol 12 myristate 13 acetate, by Merck Group (1 mentions)
Nigericin, by Cayman Chemical (1 mentions)
Polyethylenimine (pei), by Polysciences (1 mentions)
Goat anti rabbit alexa 488, by Thermo Fisher Scientific (1 mentions)
Oregon green 488 dextran, by Thermo Fisher Scientific (1 mentions)
Pbs tablet, by Thermo Fisher Scientific (1 mentions)
Texas red dextran, by Thermo Fisher Scientific (1 mentions)
A740003, by Bio-Techne (1 mentions)
3 isobutyl 1 methylxanthine ibmx, by Merck Group (1 mentions)
2 8 3h adenine, by PerkinElmer (1 mentions)
4 protocols using ionomycin
1
Fibronectin-Based Cell Activation Assay
2
Recombinant rBmpA Protein Production
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rBmpA was produced in Escherichia coli BL21 (GE Healthcare, Chicago, IL, USA) using the bacterial expression vector pGEX-6P1 (GE Healthcare) and the following primers with EcoRI and XhoI restriction sites: Forward, 5′-ACGAATTCATGAATAAAATATTGTTGTTGA-3′ and reverse, 5′-AGCTCGTAAATAAATTCTTTAAGAAA-3′ (4 (link),5 (link)). Pure ISOF was provided by Dr Weimin Yang [School of Pharmaceutical Science and Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University (KMU), Kunming, China] and reconstituted at 1×105 µM/ml in dimethyl sulfoxide (DMSO) for stock solution. LPS and phorbol-12-myristate-13-acetate (PMA) were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany) and ionomycin was purchased from Apollo Scientific Ltd. (Stockport, UK). Recombinant human (rh)IL-4, rhTNF-α and rh-granulocyte macrophage colony-stimulating factor (rhGM-CSF) were purchased from R&D Systems, Inc. (Minneapolis, MN, USA).
3
Lysosomal Trafficking and P2X Receptor Signaling
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Reagents were from Sigma-Aldrich (St. Louis, MO, USA) unless otherwise stated. Other reagents were obtained as follows: Dextran-Texas Red™ (DexTR; Mr 10,000, anionic, fixable), Dextran-Oregon Green 488 (DexOG; Mr 10,000, anionic, fixable), and LysoTracker DND 99 Red were from ThermoFisher Scientific (Paisley, UK). TransIT-LT transfection reagent was from Geneflow (Lichfield, UK), ionomycin from Apollo Scientific (Stockport, UK), polyethylenimine (PEI) from Polysciences (Warrington, PA, USA), PBS tablets from Gibco (Dublin, Ireland), Magic RedTM kit from BioRad (Hercules, CA, USA), and nigericin (sodium salt) and monensin (sodium salt) from the Cayman Chemical Company (Ann Arbor, MI, USA). Anti-P2X4 antibody (1:250; cat number APR-002; Alomone Labs, Jerusalem, Israel) and Alexa 488 goat anti-rabbit (1:250; cat number A-11008; ThermoFisher, Paisley, UK) were used in immunocytochemistry experiments. The P2X7 receptor antagonist, A740003 was obtained from Tocris (cat number 3701). Sources of additional materials are provided within the relevant methods.
4
Signaling Pathway Modulation in Cells
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N-[2-[[3-(4-bromophenyl)-2-propenyl]amino]ethyl]-5-isoquinolinesulfonamide dihydrochloride (H89) and 8-Br-cAMP were from R&D Systems (Minneapolis, MN). 2′,5′-dideoxyadenosine (DDA), N,N-dimethyl-(3R,4aR,5S,6aS,10S,10aR,10bS)-5-(acetyloxy)-3-ethenyldodecahydro-10,10b-dihydroxy-3,4a,7,7,10a-pentamethyl-1-oxo-1H-naphtho[2,1-b]pyran-6-yl ester β-alanine hydrochloride (NKH477) and 9-(tetrahydro-2-furanyl)-9H-purin-6-amine (SQ22536) were from Merck Biosciences (Middlesex, UK). [2,8-3H]-adenine was from Perkin Elmer (Waltham, MA). 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA) was from Molekula (Gillingham, UK). Carbamylcholine chloride (carbachol, CCh) and 3-isobutyl-1-methylxanthine (IBMX) were from Sigma-Aldrich (Gillingham, UK). Ionomycin was from Apollo Scientific (Stockport, UK). Cell culture media, G-418, fluo 4AM and fura 2AM were from Life Technologies (Paisley, UK). 3-[5-(tert-butyl)isoxazol-3-yl]-2-[2-(3-chlorophenyl)hydrazono]-3-oxopropanenitril (ESI-09) was from Biolog Life Science Institute (Bremen, Germany). All PTH analogues were human forms and supplied by either Bachem (Bubendorf, Switzerland) or, for PTHrP(1-36), custom-synthesized by Selleckchem (Boston, MA, USA). Sequences of the analogues used are listed in supplementary material Table S1 .
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