Mouse anti glyceraldehyde 3 phosphate dehydrogenase antibody

Tissue lysates were prepared using TissueLyser II (QIAGEN, Valencia, CA, USA) with T-PER Tissue Protein Extraction Reagent and protease inhibitor cocktail (Thermo Fisher Scientific, Waltham, MA, USA). Total protein concentration was measured by BCA Protein Assay (Thermo Fisher Scientific, Waltham, MA, USA), and equal protein concentrations were resolved by NuPAGE 4%–12% Bis-Tris SDS-PAGE (Thermo Fisher Scientific, Waltham, MA, USA). Electrophoresed proteins were transferred to nitrocellulose membranes using the iBlot Dry Blotting System (Thermo Fisher Scientific, Waltham, MA, USA) and blocked with Odyssey Blocking Buffer (PBS) (Li-Cor Biosciences, Lincoln, NE, USA). Membranes were then incubated with rabbit anti-GO, rabbit anti-LDHA (Cell Signaling Technology, Danvers, MA, USA), rabbit anti-HYPDH (Abcam, Cambridge, MA, USA), or rabbit anti-AGT (Abcam, Cambridge, MA, USA) and with mouse anti-glyceraldehyde 3-phosphate dehydrogenase antibody (Abcam, Cambridge, MA, USA). Anti-rabbit IRDye 680 and anti-mouse IRDye 800 secondary antibodies (Li-Cor Biosciences, Lincoln, NE, USA) were used for detection, and signal intensity was measured using the Odyssey Infrared Imaging System (Li-Cor Biosciences, Lincoln, NE, USA).

Tissue lysates were prepared using TissueLyser II (QIAGEN, Valencia, CA) with T-PER Tissue Protein Extraction Reagent and protease inhibitor cocktail (Thermo Fisher Scientific, Waltham, MA). Total protein concentration was measured by BCA Protein Assay (Thermo Fisher Scientific, Waltham, MA), and equal protein concentrations were resolved by NuPAGE 4%–12% Bis-Tris SDS-PAGE (Thermo Fisher Scientific, Waltham, MA). Electrophoresed proteins were transferred to nitrocellulose membranes using the iBlot Dry Blotting System (Thermo Fisher Scientific, Waltham, MA) and blocked with Odyssey Blocking Buffer (Li-Cor Biosciences, Lincoln, NE). Membranes were then incubated with rabbit anti-glycogen synthase antibody (Cell Signaling Technology, Danvers, MA) and with mouse anti-glyceraldehyde 3-phosphate dehydrogenase antibody (Abcam, Cambridge, MA). Anti-rabbit IRDye 680 and anti-mouse IRDye 800 secondary antibodies (Li-Cor Biosciences, Lincoln, NE) were used for detection, and signal intensity was measured using the Odyssey Infrared Imaging System (Li-Cor Biosciences, Lincoln, NE).