Horseradish peroxidase hrp linked secondary iggs

Foetal Bovine Serum (FBS), Eagle’s Minimum Essential Medium (MEM) and RPMI-1640 (Roswell Park Memorial Institute 1640) were obtained from Biowest (Nuaillé, France). Thin Layer Chromatography (TLC) plates and protease inhibitor cocktail were from Sigma-Aldrich (Madrid, Spain). [Methyl-¹⁴C]choline was from Perkin Elmer (Madrid, Spain). Mini-PROTEAN^® TGX Stain-Free Protein Gels, Trans-Blot Turbo Mini PVDF and Clarity Western ECL substrate were from Bio-Rad Laboratories, Inc. (Madrid, Spain). Monoclonal anti-human primary antibodies ChoKα (sc-23382) and polyclonal β-actin were from Santa Cruz Biotechnology, Inc. (Heidelberg, Germany). Rabbit polyclonal SLC44A1/CTL1 antibody (ab110767) was from Abcam (Cambridge, MA, USA). Horseradish peroxidase (HRP)-linked secondary IgGs were from Cell Signaling Technology (Danvers, MA, USA). The pET-28a vector and Escherichia coli BL21 (DE3) Star cells were from Invitrogen (Carlsbad, CA, USA), N-terminal 6x His-tag was purchased from Genescript (Piscataway, NJ, USA), Ni-NTA agarose beads were from Qiagen (Venlo, The Netherlands), and HiPrep 26/60 Sephacryl 100 HR column was from GE Healthcare (Little Chalfont, Buckinghamshire, UK). All other reagents were of analytical grade.

FBS and MEM were obtained from Biowest (Nuaillé, France). The TLC plates, protease-inhibitor cocktail, fluorimetric intracellular ROS kit, mitochondrial membrane potential kit, phosphatidylcholine assay kit, and caspase 3 assay kit were from Sigma-Aldrich (Madrid, Spain). The Muse Annexin V & Dead Cell Assay was from Merck Chemicals & Life Science (Madrid, Spain). [Methyl-¹⁴C]choline, [1,2-¹⁴C]acetic acid and [2-³H]glycerol were from Perkin Elmer (Madrid, Spain). Mini-PROTEAN® TGX Stain-Free™ Protein Gels, Trans-Blot® Turbo™ Mini PVDF and Clarity™ Western ECL substrate were from Bio-Rad Laboratories, Inc. (Madrid, Spain). Monoclonal anti-human primary antibody ChoKα and polyclonal β-actin were from Santa Cruz Biotechnology, Inc. (Heidelberg, Germany). Monoclonal anti-human primary antibodies (p62, AMPKα, Phospho-AMPKα, CHOP, IRE1α), polyclonal anti-human primary antibodies (LC3A/B and Beclin-1) and horseradish peroxidase (HRP)-linked secondary IgGs were from Cell Signaling Technology (Danvers, MA, USA). Polyclonal anti-human antibody, SREBP-2, and monoclonal anti-human antibodies LDLR and HMGCR were from Abcam (Cambridge, MA, USA).