Cibacron blue f3ga

N-isopropylacrylamide (NIPAm), N,N′ methylene bisacrylamide (BIS), allylamine (AA), potassium persulfate (KPS), vinyl sulfonic acid (VSA), Cibacron Blue F3GA (CB), dithiothreitol (DTT), iodoacetamide (IAA), urea, tris-HCl, sodium thiocynate (NaSCN), tris-(2-carboxyethyl)phosphine (TCEP), and ammonium hydroxide were procured from Sigma-Aldrich (St. Louis, MO). All solvents (water, acetonitrile [ACN], formic acid) were LC-MS grade and obtained from Fisher Scientific (Waltham, MA). Red top glass vacutainer tubes for serum separation were purchased from Becton, Dickinson and Company (Franklin Lakes, NJ). Stromal cell-derived factor 1β (SDF-1β; MW 11 kDa) was from Antigenix America (Melville, NY), insulin-like growth factor 1 (IGF1; MW 22 kDa) was from AbD Serotec (Raleigh, NC), insulin-like growth factor-binding protein 7 (IGFBP7; MW 29 kDa) was from PreproTech (Rocky Hill, NJ), and chicken lysozyme was from Sigma-Aldrich. All synthetic peptides were procured from Thermo Fisher.

During experiments, tissues were maintained in Krebs’ solution consisting of (mM): NaCl 118.4, KCl 4.7, CaCl₂ 2.5, MgSO₄ 1.2, KH₂PO₄ 1.2, NaHCO₃ 25 and glucose 11.7. ATP, tetraethylammonium chloride (TEA), tetrodotoxin, atropine sulfate monohydrate and carbachol were obtained from FUJIFILM-Wako (Osaka, Japan). D-tubocurarine, suramin, methoctramine hydrate, 4-diphenylacetoxy-N-methyl-piperidine methiodide (4-DAMP), pyridoxal phosphate-6-azophenyl-2,4-disulfonic acid (PPADS), cibacron blue F3GA (CBF3GA; synonym reactive blue 2) and 4-aminopyridine (4-AP) were obtained from Sigma-Aldrich (St Louis, MO, USA). Glibenclamide and nicorandil were obtained from Tokyo Chemical Industry (Tokyo, Japan). GSK1016790A was obtained from Cayman Chemical (Ann Arbor, MI, USA). Apamin was obtained from Peptide Institute (Osaka, Japan). tetrodotoxin was dissolved in citrate solution. Glibenclamide, nicorandil and GSK1016790A were dissolved in DMSO. Other drugs were dissolved in distilled water. The highest concentration of vehicles (0.1%) for the drugs alone had no effect on the basal tone and contractile responses at the concentrations used. The concentrations of drugs given were final concentrations in the bath solution.