Protein g sepharose 4 fast flow beads

PSA, human serum from male AB plasma and bovine serum albumin (BSA) were acquired from Sigma Aldrich. Phosphate-buffered saline (PBS) was prepared through the dilution of a PBS 10× stock solution (ThermoFisher Scientific, Waltham, MA, USA) in Milli-Q water to a final solution of 1× (pH 7.4, 25 °C). The salts, polymers, and ILs used to prepare the aqueous solutions and ABS are described in Table 1. Protein G Sepharose^® 4 fast flow beads were purchased from Cytiva, Marlborough, MA, USA. The anti-PSA mouse monoclonal capture (MABX5532) and detection (MABX5523) antibodies were acquired from Sigma Aldrich, St. Louis, MO, USA. The detector anti-PSA antibody was conjugated to the amine reactive dye Alexa Fluor^® (A430) NHS ester from ThermoFisher Scientific, Waltham, MA, USA to enable fluorescent detection. All aqueous solutions were prepared using PBS unless otherwise stated.

Human wild type (Wt), N-terminal myristoylated (myr)-tagged, and C-terminal NLS-tagged PTEN complementary DNA (cDNA) sequences were subcloned from pCDNA3-PTEN into pTRIPZ vector (Addgene, Watertown, MA, USA) with AgeI and MluI to generate PTEN expression plasmid. All plasmid constructs were confirmed by sequencing. Ampicillin, puromycin, and doxycycline were purchased from Sigma-Aldrich (St. Louis, MO, USA). Polyethylenimine (PEI) was purchased from Polysciences (Warrington, PA, USA), while RPMI, DMEM, Opti-MEM reduced serum media, and fetal bovine serum (FBS) were purchased from Thermo Fisher Scientific (Waltham, MA, USA), and Protein G Sepharose 4 Fast Flow beads were purchased from Cytiva (Malborough, MA, USA). The antibodies for Western blotting were as follows: anti-PTEN (138G6, 1:1000), anti-p-AKT (9271, 1:1000), anti-AKT (9272, 1:1000), anti-GAPDH (D16H11, 1:4000), anti-Lamin B1 (D4Q4Z, 1:1000), anti-β-Tubulin (9F3, 1:1000), and anti-EGFR (D3871, 1:1000) antibodies, all purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-HSP90 (610418, 1:4000) was purchased from BD Bioscience (Franklin Lakes, NJ, USA).