Hrp conjugated goat antirabbit igg antibodies

The protein levels of the NGAL were detected using the Western blotting analysis. Briefly, the bicinchoninic acid method was used to measure protein concentrations, and the kidney proteins (10 µg) were separated by electrophoresis in 10% sodium dodecyl sulfate polyacrylamide gels. Proteins in the gels were transferred to nitrocellulose membranes. The membranes were then probed with antiNGAL polyclonal antibodies (1:500; Bioss Biological Technology Co., LTD, Beijing, China) and antiCytc monoclonal antibodies (1: 2000; Bioss Biological Technology Co.) for rabbit proteins. HRP-conjugated goat antirabbit IgG antibodies (Cell Signaling Technology, Shanghai, China) were applied as secondary antibodies (1: 10,000). The relevant protein bands were visualized by enhanced chemiluminescence reagents and analyzed with the Odyssey image analysis system. To determine an equal content of proteins for each sample, the membranes were reprobed with mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) polyclonal antibodies (1: 5000, Cell Signaling Technology). The result was expressed as the ratio of target protein to GAPDH.

Rabbit monoclonal anti-IRF3, GAPDH, eGFP-Tag, HA-Tag and HRP-conjugated goat anti-rabbit IgG antibodies, and mouse monoclonal anti-Myc-Tag Flag-Tag antibody were purchased from Cell Signaling Technology (USA). The JetPRIME kit was purchased from Polyplus Transfection (France) and Double-Luciferase Reporter Assay Kit was purchased from TransGen (China). HRP-conjugated goat anti-mouse-IgG antibody, protease inhibitor, and phosphatase inhibitor were provided from CWBIO (China). The Pierce Crosslink Magnetic IP/Co-IP Kit was purchased from Thermo Scientific (USA). Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) and Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) were purchased from Abcam (USA). TRIM21 Polyclonal antibody was purchased from Proteintech (USA). MG132, chloroquine diphosphate (CQ), 3-methyladenine (3-MA) and Z-VAD-FMK were purchased from MedChemExpress (MCE) (USA).