Secondary goat anti rabbit igg

The proteins were loaded onto Novex Tris-Glycine 4–20% gels (Life Technologies EC6025) and then transferred overnight onto nitrocellulose paper (Bio-Rad Laboratories, cat. # 262–0115). Membranes were blocked with Li-COR Odyssey Blocking Buffer (Li-COR Biosciences, Lincoln, NE, cat. # 927–4000) diluted in Tris Buffered Saline (TBS). Antibodies from Abcam (Cambridge, UK) were anti-SDHB (SDH) [cat. # ab14714], mitoProfile pyruvate dehydrogenase WB Antibody Cocktail (PDHC), anti-pyruvate dehydrogenase (PDH) cocktail (cat. # ab110416) for the following proteins: 69 kDa E2, 54 kDa E3, 43.3 kDa E1[alpha], 39.4 kDa E1[beta], anti-aconitase 2 (cat. # ab71440), anti-MDH2 (MDH, cat. # ab96193) and antiICDH (cat. # ab113232). STK19 primary antibody (STH, cat. # Sc-28027) was purchased from Santa Cruz Biotechnology (Dallas, TX). Primary β-actin rabbit monoclonal antibody from Cell Signaling Technology (Beverly, MA) or mouse β-actin primary polyclonal antibody (Cell Signaling, cat. # 4970S) were added to detect β-actin. Secondary goat anti-rabbit IgG (Li-COR, cat. # 926–68071) and secondary goat anti-mouse antibodies (Li-COR, cat. # 926–32210), were added to detect actin and the protein of interest. The membranes were scanned using the LiCOR Odyssey instrument.