Goat anti rabbit igg h l hrp antibody

The total protein was extracted using RIPA lysis buffer (Thermo Fisher Scientific, Waltham, MA, USA) with protease inhibitor and phosphatase inhibitors (Beyotime, Nanjing, China) and quantified by a BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA, USA). Total protein extracts (30 µg) from exosomes, cells, and submandibular gland tissue were separated via 10% SDS‑PAGE (Epizyme, Shanghai, China) and transferred to polyvinylidene fluoride membranes (Millipore, Burlington, MA, USA) and blocked with 5% non‑fat milk for 1 h at room temperature. After incubation with the primary antibody and the secondary antibody, the target protein was visualized by chemiluminescence using an ECL kit (Beyotime, Nanjing, China). The antibodies used in the Western blot assay are listed as follows: GAPDH (1:10,000, ab181602, Abcam), AQP5 (1:1000, sc-514022, Santa Cruz), CD63 (1:1000, ab134045, Abcam), TSG101 (1:1000, ab125011, Abcam), ALIX (1:1000, ab275377, Abcam), GPER (1:1000, ab260033, Abcam), CREB (1:1000, ab32515, Abcam), p-CREB (1:5000, ab32096, Abcam), PKA (1:1000, #4782, Cell Signaling Technology), p-PKA (1:1000, #4781, Cell Signaling Technology) and goat anti-rabbit IgG H&L (HRP) antibody (1:5000, #31460, Thermo Fisher Scientific).

The in-house guinea pig anti-MERS-CoV nucleocapsid protein (NP) serum and negative control guinea pig serum were used for MERS-CoV NP detection in flow cytometry experiments as we previously described [16 (link)]. Mouse anti-hnRNP C antibody (clone 4F4, IgG1) (Santa Cruz Biotechnology, sc-32308) was used in flow cytometry, Western blot, and RNA immunoprecipitation (RIP) assay. Anti-eIF4EBP1 (phosphor T36) antibody (Abcam, ab47365), anti-phospho-hnRNP C (Ser260) antibody (Thermo Fisher Scientific, Waltham, MA, USA, PA5-38589), anti-mTOR antibody (Abcam, ab134903), anti-mTOR (phospho S2481) antibody (Abcam, ab137133), anti-CRK antibody (Abcam, ab133581), anti-β-actin antibody (Sigma, A5441) were used in western blot analyses as primary antibodies for the detection of each respective target protein. Goat anti-rabbit IgG (H + L) HRP antibody (Thermo Fisher Scientific, 656120), and goat anti-mouse IgG (H + L) HRP antibody (Thermo Fisher Scientific, 626520) were used as secondary antibodies in the western blot. Rabbit IgG (Abcam, ab37415) and mouse IgG1 (Abcam, ab81032) were used as isotype control antibodies.