Chemikine nerve growth factor sandwich elisa kit

In order to prevent any release of loosely bound factors NGF and CNTF, functionalized electrospun nanofibers (n = 3) were incubated in 2 mL of sterile PBS containing 0.1% BSA at 37°C without any pre-treatment (like ethanol sterilization) and 150 µL were harvested after 1, 2, 8, 48 and 120 hours and stored at −80°C. NGF and CNTF release assays were performed with the ChemiKine Nerve Growth Factor Sandwich ELISA Kit (Millipore, France) and Raybio Rat CNTF ELISA kit (TEBU-BIO, France) respectively, according to the manufacturer's instructions.

Poly(ε-caprolactone) (M_w 80 kDa), glutaraldehyde, sorbitan monooleate (Span 80), hexamethyldisilazane (HMDS), sodium dodecyl sulfate (SDS), Dulbecco’s modified eagle’s medium (DMEM/F12) were purchased from Sigma, Singapore. Chloroform (CHCl₃) was purchased from Fisher Scientific Company, Loughborough, UK. Micro BCA Protein Assay Kit was purchased from Thermo Scientific (Rockford, lL, USA). Sodium hydroxide (NaOH) was bought from Merck (Singapore). Nerve growth factor (NGF 7S) and ChemiKine Nerve Growth Factor Sandwich ELISA kit were purchased from Millipore, Singapore. Rat pheochromocytoma (PC12) cells in the adherent type [PC12 Adh (ATCC^® CRL1721.1™)] were obtained from ATCC, Manassas, VA, USA, while fetal bovine serum (FBS), horse serum (HS), and trypsin/EDTA were purchased from Invitrogen/Gibco, Grand Island, NY, USA. [3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt; MTS] was purchased from Promega, Madison, WI, USA. Alexa Fluor 594 Goat anti-Mouse IgG antibody was obtained from Invitrogen, Eugene, OR, USA.