2 mercaptoethanol

GO was purchased from Graphene Supermarket (Calverton, NY, USA) and 6-arm poly(ethylene glycol)-amine (15 kDa) was purchased from SunBio (Seoul, Korea). N-3-(dimethylamino)propyl-N′-ethylcarbodiimide hydrochloride (EDC) and chloroacetic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA). 2-mercaptoethanol and sodium hydroxide were purchased from Bio Basic Inc (Ontario, Canada). 10× isothermal amplification buffer II, 100 mM MgSO₄, and Bst 3.0 DNA polymerase were purchased from New England Biolabs (Ipswich, MA, USA). Deoxynucleotides (dATP, dTTP, dGTP, dCTP: 100 mM each), and SYBR^™ Green I were purchased from Invitrogen (Carlsbad, CA, USA). GelRed^® nucleic acid gel stain was purchased from Biotium (Fremont, CA, USA).

Cells were suspended in SDS protein sample buffer ((140 mM Tris (EMD, catalogue number 9230) pH 6.8, 4% SDS (BioShop, catalogue number SDS001), 20% glycerol (BioShop, catalogue number Gly002.4), 0.02% bromophenol blue (BioShop, catalogue number BRO222.5), 1:100 diluted 2-mercaptoethanol (Bio Basic Canada, catalogue number MB0338) and incubated on ice for 15 min. Lysates were boiled for 5 min and clarified in a microfuge at 15 000 g for 2 min at 4ºC. The supernatant was run on a 10% SDS polyacrylamide gel and transferred to an activated PVDF membranes (Bio-Rad, catalogue number 162–0177) in a Tris-glycine transfer buffer containing 20% methanol (Caledon Laboratory Chemicals, catalogue number 6701-7-40) in a Gel Transfer Cell (BioRad catalogue number 1703930). Horseradish peroxidase-conjugated goat anti-mouse IgG (H + L) (Thermo Fisher Scientific, catalogue number 31430) or anti-rabbit IgG (H + L) (Thermo Fisher Scientific, catalogue number 31460) secondary antibodies were used at a dilution of 1:5000. Blots were developed using the ECL Western Blotting Detection Reagent (Cytiva Amersham, catalogue number RPN2106) and visualized by using MicroChemi 4.2 (Bio-Imaging Systems).