
Western blot analysis of extracts from serum starved HeLa cells, untreated (-) or treated with insulin (150 nM, 5 min; +) or IGF-1 (100 ng/ml, 5 min; +), using Phospho-PFKFB2 (Ser483) (D4R1W) Rabbit mAb (upper) or PFKFB2 (D7G5R) Rabbit mAb #13045 (lower). The phospho-specificity of Phospho-PFKFB2 (Ser483) (D4R1W) Rabbit mAb is verified by treating the nitrocellulose membranes with calf intestinal alkaline phosphatase (CIP) and λ protein phosphatase (λ PP) following transfer of proteins from the SDS-PAGE gel to the membranes.