Anti mouse igg dylight680

The primary antibodies: anti-Cyclophilin A (1:1000 dilution, Abcam, ab126738), anti-CD147 (1:1000, ThermoFisher, MA1–19,201), anti-HSP70 (1:3000, Santa Cruz, sc-66,048), anti-TSG101 (1:1000 dilution, Abcam, ab125011), anti-HSP90B1 (1:1000 dilution, Cusabio, CSB-PA10887A0Rb), anti-GAPDH (1:500 dilution, EMD Millipore, MAB374), anti-Rab7 (1:1000 dilution, Cell signaling Technology, 9367), anti-Rab11 (1:250 dilution, ThermoFisher, 71–5300).
The secondary antibodies: anti-Rabbit IgG-DyLight 800 (1:10,000 dilution, ThermoFisher, SA5–35,571), anti-Mouse IgG-DyLight 680 (1:10,000 dilution, ThermoFisher, 35,519)
Reagents: Trypsin (Pierce Trypsin Protease, MS Grade, ThermoFisher, 90,057), Propidium Iodide (ThermoFisher, P1304MP), 5-(and −6)-Carboxyfluorescein Diacetate, Succinimidyl Ester, mixed isomers (5(6)-CFDA, SE) (ThermoFisher, C1157)

Protein expression was assessed by Western blotting. Cells were harvested from the culture plates and total cellular proteins were extracted by lysis buffer containing 1% Triton X-100, 1% proteasome inhibitor cocktail (Sigma-Aldrich). Lysates were cleared by centrifugation at 13,000 g for 20 min and protein concentrations were measured using the BC Assay Protein Quantitation Kit (Uptima, Oakland, CA). Total protein lysates (50 μg) were fractionated on 15% SDS-PAGE then transferred to nitrocellulose membranes (GE Healthcare Life Sciences, Vélizy-Villacoublay, France). Membranes were blocked for 1 h in PBST with 2.5% BSA, and then incubated with primary antibodies overnight at 4 °C, rabbit polyclonal anti-BDNF (sc-546, Santa Cruz [51 (link)]) at 1:200 and mouse monoclonal anti-α-tubulin (Clone DM1A, Sigma-Aldrich) at 1:10,000. Incubation with secondary antibodies conjugated to infrared fluorophores (goat anti-rabbit IgG Dylight 800 and anti-mouse IgG Dylight 680 at 1:10,000 from Thermo Scientific) was performed for 1 h. An Odyssey infrared imaging system (LI-COR, Bad Homburg, Germany) was used to scan membranes at a wavelength of 680 nm (anti-mouse) or 800 nm (anti-rabbit). Data were analyzed with Image Studio 1.1 software (Li-COR).