Cl 488 conjugated affinipure goat anti rabbit lgg h l

The LLC‐SD‐siN.C. and LLC‐SD‐siNr5a2 (LLC‐SD‐siNr5a2‐1/LLC‐SD‐siNr5a2‐2) cells were analyzed for CD133 cell‐surface marker expression by flow cytometry according to the manufacturer's protocol. The primary antibody was used at the following working dilutions: anti‐CD133 antibody (0.2 μg 18470‐1‐AP, Proteintech, USA), and the secondary antibody: CL‐488‐conjugated Affinipure Goat Anti‐Rabbit lgG (H + L) (1:100‐1:500 dilution, Proteintech, USA).

Berberine chloride dihydrate (Berberine hydrochloride, purity: 98%) was obtained from Yuanye Inc. (Shanghai, China) and Ibuprofen (purity: 98%) was obtained from Aladdin (Shanghai, China). Both of them were used directly after purchase. All other solvents were analytically pure. Deionized water was used in this study. All plasmids were supplied by Shanghai GenePharma Co. Ltd (Shanghai, China). Sources of antibodies were as follows: IKKε antibody (Cell SignalingTechnology, 2690), TBK1 antibody (Cell Signaling Technology, 3013), Phospho-TBK1 Ser172 antibody (Cell Signaling Technology, 5483), Phospho-AMPKα Thr172 antibody (Cell Signaling Technology, 2535), Anti-UCP1 antibody (Abcam, ab10983), F4/80 antibody (proteintech, 28463-1-AP), GAPDH antibody (proteintech, 60004-1-Ig), Anti-rabbit IgG (Cell Signaling Technology, 7074 S), Anti-mouse IgG (Cell Signaling Technology, 7076 S), CL488-conjugated Affinipure Goat Anti-Mouse lgG (H + L) (proteintech, SA00013-1), CL488-conjugated Affinipure Goat Anti-Rabbit lgG (H + L) (proteintech, SA00013-2). 3-Isobutyl-1-methylxanthine (IBMX), dexamethasone (DEX), insulin, ISO and Oil Red O solution (0.5% in isopropanol) were purchased from Sigma-Aldrich (St Louis, MO, USA). Recombinant Murine TNF-α was purchased from Pepro Tech (Rocky Hill, NJ, USA). The BCA protein assay kit was purchased from Beyotime Biotechnology Co., Ltd (Beijing, China).