Confocal immunofluorescent analysis of LNCaP (high expression, left) and HepG2 (low expression, right) using PFKFB2 (D5I5F) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
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.
Immunoprecipitation of PFKFB2 from Hep G2 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or PFKFB2 (D5I5F) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using PFKFB2 (D5I5F) Rabbit mAb. An anti-rabbit IgG light chain antibody was used as the secondary antibody.
Western blot analysis of extracts from LNCaP, Hep G2, and MCF7 cells using PFKFB2 (D5I5F) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.
The bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK/FBPase or PFKFB) catalyzes the synthesis and degradation of fructose 2,6-bisphosphate and regulates its steady-state level (1,2). Fructose 2,6-bisphosphate activates phosphofructokinase, a rate-limiting enzyme in glycolysis, by allosteric regulation (1,2). Four different PFKFB isoforms (PFKFB1, PFKFB2, PFKFB3, and PFKFB4) have been identified (1,2). Research studies indicate that amino acids activate PFKFB2 through Akt-dependent phosphorylation at Ser483 on PFKFB2 (3). In addition, androgen increases the expression of PFKFB2 in prostate cancer cells (4).
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