Rabbit polyclonal anti lc3b antibody

Most of the chemicals were obtained from Sigma (St. Louis, MO, USA). Structurally defined, novel, and pure (>98.9% purity) garcinielliptone FC [7 (link)] and justicidin A [8 (link)] were gifts from Dr. Chun-Nan Lin (School of Pharmacy, Kaohsiung Medical University, Kaohsiung, Taiwan) and formosanin C (FC) was kindly donated by Dr. Shen-Jeu Won (College of Medicine, National Cheng Kung University, Tainan, Taiwan) [9 (link)]. Sorafenib (Nexavar^®) was purchased from Bayer AG (Leverkusen, Germany). C11-BODIPY was obtained from Thermo Fisher Scientific Inc. (Waltham, MA, USA). Antibodies for Western blotting were purchased from the following vendors: rabbit monoclonal anti-CD71 (TFRC) and anti-ferritin heavy chain 1 (FTH1) antibodies, Cell Signaling Technology, Danvers, MA, USA; rabbit polyclonal anti-FPN/SLC40A1 antibody, Novus Biologicals, LLC, Littleton, CO, USA; rabbit polyclonal anti-ARA70 (NCOA4) antibody, Bethyl Laboratories, Inc., Montgomery, TX, USA; rabbit polyclonal anti-LC3B antibody, Abcam, Cambridge Science Park, Cambridge, UK; rabbit polyclonal anti-GAPDH antibody, GeneTex, Irvine, CA, USA; mouse monoclonal anti-β-actin antibody, Sigma; goat anti-rabbit (H+L) and goat anti-mouse IgM+IgG+IgA (H+L) HRP conjugate antibodies, Millipore Corp., Billerica, MA, USA.

Grass carp LC3B was subcloned into pEGFP-N3 (Clontech, Mountain View, CA, USA) to generate GFP-LC3B to detect the formation of autophagy. Grass carp ATG5 was subcloned into pCMV-FLAG, pDsRed2-C1, and pEGFP-N3 (Clontech, Mountain View, CA, USA ) to generate FLAG-ATG5, DsRed-ATG5, and GFP-ATG5 to overexpress ATG5 in CIK cells. The plasmids were constructed as described previously [30 (link),31 (link)] Then, these vectors (500 ng/µL) were transfected into CIK cells by Lipofectamine™ 3000 Transfection Reagent (Invitrogen, Carlsbad, CA, USA), according to the manufacturer’s recommendations. Rapamycin, 3-MA, and horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG were purchased from Sigma (Saint Louis, MS, USA). Rabbit polyclonal anti-LC3B antibody was purchased from Abcam (Cambridge, UK). Rabbit polyclonal SQSTM1/P62 antibody was purchased from Beyotime (Shanghai, China). Lyso-Tracker Red fluorescent probe was purchased from Solarbio (Beijing, China).