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2 protocols using tat peptide

1

Protein Conjugation and Cellular Uptake Assay

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Donepezil, clioquinol, HSA, ThT, 1‐(3‐dimethylaminopropyl)‐3‐ethylcarbodiimide hydrochloride (EDC), N‐hydroxysuccinimide (1‐hydroxypyrrolidine‐2,5‐dione, NHS), and FITC‐dextran were purchased from Sigma–Aldrich; GM1 was purchased from J&K(Beijing); TAT peptide and Aβ1‐42 were purchased from GL Biochem (Shanghai), and the purity of the peptides was greater than 98%; glutaric dialdehyde was purchased from Aladdin (Shanghai); hexafluoroisopropanol (HFIP) and 4‐(N‐maleimidomethyl) cyclohexane‐1‐carboxylic acid 3‐sulfo‐N‐hydroxysuccinimide ester sodium salt (Sulfo‐SMCC) were purchased from Yuanye (Shanghai). FITC‐transferrin, FITC‐cholera toxin subunit B, and LysoTracker Red probe were purchased from Invitrogen (USA). Milli‐Q water (18 MΩ cm) was used in the experiments. Nicotinic acetylcholine receptors α7(α7 nAChR), SYAP1 and GAP43 antibody were purchased from Bioss (Beijing). Rat cysteine protease‐3 ELISA kit was purchased from Huyu Biotechnology (Shanghai).
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2

Synthesis and Characterization of TAT-Functionalized Gold Nanoparticles

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Chloroauric acid (HAuCl4) was purchased from Civi-Chem (Suzhou, China). HS-PEG-COOH (MW = 5,000) were purchased from Pengshuo Biotech Co., Ltd. (Shanghai, China). Silver nitrate (AgNO3) was purchased from Alfa (America). Ascorbic acid (AA) was purchased from Macklin (Shanghai, China). Hydrochloric acid (HCl) was purchased from Sinopharm Chemical Reagent Co. Ltd (Shanghai, China).1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-Hydroxysuccinimide (NHS) were purchased from Aladdin (Shanghai, China). Phosphate buffer saline (PBS), high glucose Dulbecco's Modified Eagle's Medium (DMEM) were purchased from HyClone (America). Penicillin/ streptomycin and fetal bovine serum (FBS) were purchased from Gibco (America). Basic fibroblast growth factor (bFGF) was purchased from Invitrogen (America). TAT-peptide (residues 47-57 sequence YGRKKRRQRRR-NH2) and FITC labeled TAT were purchased from GL Biochem Ltd (Shanghai, China). Cell counting kit-8 (CCK-8) was purchased from Beijing Zoman Biotechnology Co., Ltd. Propidium Iodide (PI) stain, 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI), Western and IP kit was purchased from Beyotime Biotechnology (Shanghai, China). Culture-Inserts were purchased from Ibidi (Germany).
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