3 mercaptopropyl triethoxysilane

Template (Pluronic P-123), 1-butanol (99.8%), camphene (98%), nonane (99%), tetraethylorthosilicate (TEOS), HCl (37%), tungstophosphoric acid, (3-mercaptopropyl) triethoxy-silane, hydrogen peroxide (30%), ethanol (96%), and acetone (99%) were acquired from Sigma–Aldrich.

All chemicals were of analytical grade and were used as received, unless otherwise specified. All aqueous solutions were made with double de-ionised water, 18.2 MΩ cm, with a Pyrogard filter (Millipore, USA). Tris base, MgCl₂, NaCl, acetone, NH₄OH, HCl, H₂O₂, 3-glycidoxypropyltrimethoxysilane (GOPTS), PKC-α kinase inhibitor GF 109203X, adenosine tri-phosphate (ATP), adenosine 5'-[γ-thio] triphosphate (ATP-S), 3-mercaptopropyl triethoxysilane (MPTES), 3,3′,5,5′-tetramentylbenzidine (TMB), allyl hydroxyl silane (AHS) and horseradish peroxidase (HRP) were purchased from Sigma-Aldrich. Dephosphorylated myelin basic protein (MBP), purified from bovine brain using FPLC liquid chromatography, was purchased from Millipore. Protein Kinase C alpha (PKC-α) human recombinant kinase produced in Sf9, was procured from ProSec-Tray TechnoGene Ltd. PKC lipid activator cocktail was obtained from Millipore. Polyclonal phospho-(Ser) PKC substrate antibody was purchased from Cell Signaling Technology. 20 nm research grade gold nanoparticles (AuNPs) were bought in colloidal form from Diagnostic Consulting Network.

We used NTA-Ni²⁺ functionalized coverslips as substrates for SMFS to bind full-length ectodomains from the tip-link cadherins, as well as pFS-1 polyproteins, via their C- and N-terminal Histags, respectively. Coverslips were first immersed into a 20 N KOH solution overnight, after which they were transferred to a 2% 3-(mercaptopropyl)triethoxysilane (Sigma-Aldrich), 0.02% acetic acid at 90 °C for 1 h. Afterwards, they were washed in a MilliQ water flow for 1 h and then cured for 15 min in an oven at 120 °C. Next, they were transferred to a 100 mM DTT solution for 15 min and washed under a MilliQ water flow for 1 h. Then, 50 μl of a solution of 20 mg/ml maleimide-C 3-NTA (Dojindo Laboratories) dissolved in 10 mM 3-(N-morpholino)propanesulfonic acid (pH 7) were added and the substrates were incubated for 1/2 h. After a final wash in MilliQ water, a drop of 10 mM NiCl₂ was added (>50 μl) on the coverslips, incubated for 10 min, and then they were washed with MilliQ water before storage.

MCM-41 hexagonal structured silica, 36.5–38 wt.% hydrochloric acid, poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol) (Pluronic P123, average molecular weight 5800), poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol) (Pluronic F127, average molecular weight 12,600), potassium hydroxide, 98% sulfuric acid, sodium chloride, 1,3,5-trimethylbenzene (TMB), toluene, ethanol were purchased from Sigma-Aldrich (Merck Group, Darmstadt, Germany), 25 wt.% ammonia aqueous solution from Scharlau (Scharlab S.L., Barcelona, Spain), trimethylhexadecylammonium bromide (CTAB, Alfa Aesar, Ward Hill, MA, USA), and tetraethyl orthosilicate, (TEOS, Fluka, Seelzer, Germany). All organosilanes: (3-aminopropyl)triethoxysilane (APTES, 95%), (2-cyanoethyl)triethoxysilane (CETES, 97%), (3-isocyanatopropyl)triethoxysilane (ICPTES, 95%), triethoxyphenylsilane (TEPS, 98%), (3-mercaptopropyl)triethoxysilane (MPTES, 95%), and toluene (anhydrous, 99.8%) were acquired from Sigma-Aldrich and used without further purification. Resveratrol (≥99% purity) was purchased from Evolva SA (Reinach, Switzerland), and ultrapure deionized water (Millipore Direct-Q3UV with Biopack UF cartridge, Merck Group, Darmstadt, Germany) was used in all experiments.

Tetraethyl orthosilicate (TEOS, ≥ 99%), cetyltrimethylammonium bromide (CTAB, ≥ 99%), (3-aminopropyl)triethoxysilane (APTES, 99%), bis[3-(triethoxysilyl)propyl] tetrasulfide (TESPTS, ≥ 90%), (3-mercaptopropyl)triethoxysilane (MPTES, ≥ 80%), acid Fuchsin (AF, dye content 70%), acid orange II (AO, ≥ 98%) were purchased from Sigma-Aldrich (Germany). Cyclohexane (≥ 99.9%), hydrochloric acid (HCl, 37%), urea (≥ 99%), n-amyl alcohol (≥ 98.5), and sodium hydroxide (NaOH, pellets, ≥ 97%) were purchased from Merck Millipore (Germany). Ethanol (96% and absolute) and acetone (HPLC grade) were purchased from Mojallali Chemical Co. (Tehran, Iran).