Dylight 488 n hydroxysuccinimide nhs ester

AZD9291, gefitinib, afatinib, and crizotinib were purchased from Selleckchem. SHR‐A1403, the naked anti‐c‐Met monoclonal antibody c‐Met mAb and free toxin SHR152852, were provided by Jiangsu Hengrui Medicine Co. Ltd.31 DyLight 488 N‐hydroxysuccinimide (NHS) ester was purchased from Thermo‐Fisher Scientific. Sulforhodamine B was purchased from Sigma‐Aldrich.
Antibodies against EGFR, phospho‐EGFR (Tyr1173), c‐Met, phospho‐c‐Met (Tyr1234/1235), STAT3, phospho‐STAT3 (Tyr705), AKT, phospho‐AKT (Ser473), ERK1/2, phospho‐ERK1/2 (Thr202/Tyr204), and GAPDH were purchased from Cell Signaling. β‐Tubulin antibody was purchased from Sigma‐Aldrich.

One milligram of purified MaSp2 N-R6-C or N-R12-C was buffer-exchanged against a solution containing 20 mM sodium phosphate buffer (pH 8.5) and 150 mM NaCl using an Amicon Ultra [50-kDa molecular weight cutoff (MWCO)] and then reacted overnight at 4°C with 50 μg of DyLight 488 N-hydroxysuccinimide (NHS) ester (Thermo Fisher Scientific), which was followed by further centrifugation in an Amicon Ultra (50-kDa MWCO) to obtain concentrated fluorescently labeled protein samples. No differences in phase separation behavior or mesoscale structural morphology were observed between the unlabeled and labeled samples.