N hydroxysuccinimide nhs

Cocoons of Bombyx mori (silkworm) were procured from farmers in Xuzhou, Jiangsu. Cs powders (900 000 Da, 95% deacetylated) were purchased from Shanghai Macklin Biochemical Technology Co. Ltd. Alg, sodium carbonate, lithium bromide, acetic acid, aqueous ethanol, polyethylene glycol 6000, dimethyl sulfoxide, cell counting kit (CCK)-8, 4% paraformaldehyde, hematoxylin, eosin, crystal violet, and dialysis bags were obtained from Shanghai Yuanye Biotechnology Co. Ltd. Ethylene dichloride (EDC) and N-hydroxysuccinimide (NHS) were bought from Sangon Biotech (Shanghai) Co. Ltd. Minimum essential medium culture media, fetal bovine serum, and penicillin–streptomycin were bought from Thermo Fisher Scientific Inc. (Waltham, MA). Four-week-old female BALB/c-nu mice were purchased from the Animal Center of East China Normal University. Mice were bred and maintained under specific-pathogen-free conditions. The study was approved by the Hospital Ethical Committee (2019 JS-004, Shanghai Fifth People’s Hospital), and all methods used in this study were carried out following the Regulations for the Administration of Affairs Concerning Experimental Animals approved by the State Council of the People’s Republic of China. Commercially available human CC cells HCT-116 (1×10⁷ cells/0.1 mL per site) were injected subcutaneously in the left underarm and right inguinal area of mice.

ssDNA library for SELEX and primers were synthesized by Sangon Biotech (Shanghai China), bovine serum albumin (BSA, Sigma-Aldrich, Japan, Catalogue#:V900933), DMEM/F12 medium (Gibco, US, Catalogue#: 12400-024), Fetal bovine serum (Gibco, US, Catalogue#: 16000-044), trypsin (Sigma-Aldrich, Japan, Catalogue#: T2600000), cell counting kit 8 (CCK8, Sigma-Aldrich, US, Catalogue#:96992), Rat osteopontin (OPN) ELISA kit (Cusabio, Wuhan, China, Catalogue#: CSB-E08392h), osteocalcin (BGP) ELISA kit (Sigma-Aldrich Co. LLC., Shanghai, China), Alkaline phosphatase (ALP) (Sigma-Aldrich Co. LLC), streptavidin-coated magnetic beads (Promega, Madison, WI), 1-ethyl-3-(3-dimethylaminopropyl)-car-bodiimide hydrochloride (EDC, Sangon Biotech, Shanghai, China, Catalogue#:C600433), N-Hydroxysuccinimide (NHS, Sangon Biotech, Shanghai, China, Catologue#:C100219).