Sulfo cyanine 5 cy5 nhs ester

Texin 985A is a polyether-based thermoplastic polyurethane (PU) obtained from Covestro, Leverkusen, Germany. N, N-dimethyl formamide (DMF, >99.8%), tetraethylorthosilicate (TEOS, >99%), (3-aminopropyl)triethoxysilane (APTES, 99%), dimethylsulphoxide (DMSO, >99%), hexamethyldisilazane (HMDS, 99.9%), bovine serum albumin (BSA; >98% heat shock fraction, protease-free, fatty acid-free, essentially globulin free, pH 7), sodium azide (99%), ethylenediaminetetraacetic acid (EDTA), paraformaldehyde (PFA), Triton-X, and 4′,6-diamidino-2-phenylindole (DAPI) were obtained from Sigma Aldrich, Darmstadt, Germany. Sulfo-Cyanine 5 (Cy5) NHS ester was bought from Lumiprobe, Germany. Absolute ethanol (>99.8%) was purchased from VWR, Dietikon, Switzerland, and ammonia solution (25%) was purchased from Merck, Buchs, Switzerland. Phalloidin-AlexaFluor 488 (Cat #A12379) and secondary antibody goat anti-mouse AF647 (Cat #A21235) were bought from Invitrogen, Thermo Fisher Scientific, Basel, Switzerland. Transforming growth factor beta (TGF-β, Cat #11343160) and tumor necrosis factor alpha (TNF-α, Cat #11343015) were acquired from Immunotools, Friesoythe, Germany. All cell culture materials were purchased from Gibco, Thermo Fisher Scientific, Basel, Switzerland, unless otherwise specified. This study made use of only Milli-Q water.

Ten molar excess of Sulfo-Cyanine5 (Cy5) NHS ester (Lumiprobe) was dissolved in 25 μL of dimethyl sulfoxide solution, and diluted to 100 μL with water. Nine Hundred micro liter of recombinant PA protein (1–2 mg/ml) in 50 mM sodium phosphate buffer pH 8 was added to the dye solution, vortexed, and stored at 4°C overnight. The unreacted dye was removed using a 10K MWCO Amicon Ultra-4 Centrifugal Filter (Merck).