Gpcmax ve 2001 gpc solvent sample module

¹H NMR and ¹³C NMR spectroscopy were performed on Avance DRX 400 MHz. Dimethylformamide (DMF) GPC was conducted in DMF containing 0.10 M LiBr (40 °C, 0.8 mL/min) on a Shimadzu HPLC system equipped with a refractive index detector RID-10A, one Polymer Laboratories PLgel guard column, and two Polymer Laboratories PLgel 5 μm mixed D columns. Calibration was performed using near-monodisperse polystyrene standards. Chromato-grams were processed using the EZStart 7.2 chromatography software. Aqueous GPC was performed in 0.05 M Na₂SO₄ in MQ water with 10% MeOH on a Viscotek GPC_max VE 2001 GPC solvent/sample module equipped with Viscotek TDA 305 triple array detector. Gel electrophoresis was performed using Any kDTM Mini-PROTEAN TGXTM precast gels with SDS-Tris-glycine as running buffer (Biorad, Hercules). ELISA assays were read on an ELX800 Universal Microplate Reader (Bio-Tek Instrument Inc., Winooski) with λ = 450 nm for signal and 630 nm for background. Western blot was developed on a FluorChem FC2 System version 3.2 (Cell Biosciences, Santa Clara). CellTiter-Blue assays were read on a SpectraMax M5 microplate reader (Molecular Devices, Sunnyvale).

PDA-PEG-PNiPMA polymer was synthesized by free radical polymerization as shown in Figure 1A. PDA monomer was prepared following our previously reported method [14 (link)]. Typically, PDA (241.3 mg, 1 mmol), Poly(ethylene glycol)methacrylate (Mn = 360 Da) (360 mg, 1 mmol) and N-isopropylmethacrylamide (NiPMA, 63.6 mg, 0.5 mmol) were dissolved in 10 mL degassed anisole. 2, 2-azobisisobutyronitrile (AIBN, 14 mg, 0.085 mmol) in 1 mL degassed anisole was then added, and the reaction mixture was stirred for 24 hours at 65 °C. The final product was precipitated (3×) in ice cold ether and dried for 48 hours in vacuum. The structure of PDA-PEG-PNiPMA was confirmed by ¹H-NMR (Figure S1A). Gel permeation chromatography (viscotek GPCmax VE 2001 GPC solvent/sample module, Viscotek VE 3580 RI detector and 270 Dual Detector) using THF as mobile phase was empolyed to charactrize the polymer and found PDA-PEG-PNiPMA has an absolute molecular weight of 27,557 Da (Mw) and polydispersity (PDI: 1.35) (Figure S1B). For the quantification of side chain functionality, PDA-PEG-PNiPMA (1.0 mg/mL) was dissolved in DMSO and incubated with dithiothreitol (DTT, 10 mM) for 1 hour at room temperature, and then the amount of 2-pyridinethione released was quantified through UV-Vis spectrophotometer at λ = 375 nm (ε, molar absorption coefficient = 8080 M⁻¹cm⁻¹).