PFKFB2 Antibody [E19B24] | Primary Antibodies

Application: Reactivity: Human, Monkey

Lane 1: LNCAP, Lane 2: HepG2, Lane 3: MCF7

Usage Information

Dilution
WB1:1000 IP1:50 IF1:1600

Application
WB, IP, IF

Reactivity
Human, Monkey

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
55 kDa

Positive Control	LNCaP cells; MCF7 cells
Negative Control

Datasheet & SDS

Biological Description

Specificity
PFKFB2 Antibody [E19B24] detects endogenous levels of total PFKFB2 protein.

Clone
E19B24

Synonym(s)
PFKFB2; 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 2; 6PF-2-K/Fru-2,6-P2ase 2; PFK/FBPase 2; 6PF-2-K/Fru-2,6-P2ase heart-type isozyme

Background
PFKFB2, the heart and muscle-predominant bifunctional enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase, precisely regulates glycolytic flux by controlling levels of fructose-2,6-bisphosphate, the most potent allosteric activator of phosphofructo-1-kinase, thereby accelerating glycolysis and inhibiting gluconeogenesis, a role especially vital in cardiac energy metabolism under stress, insulin, and adrenergic regulation. The enzyme forms a head-to-head homodimer, with each monomer containing an N-terminal regulatory segment, a central kinase domain with bi-lobed adenylate kinase-like architecture featuring a six-stranded β-sheet and α-helices, and an ATP/fructose-6-phosphate binding pocket with key catalytic residues that coordinate Mg²⁺-ATP. Its C-terminal phosphatase domain, homologous to histidine phosphatases, contains flexible loops that grant fructose-2,6-bisphosphate access for hydrolysis via a phosphohistidine relay, while the C-terminal tail harbors Ser466/483 sites for Akt and PKA phosphorylation. The kinase activity synthesizes fructose-2,6-bisphosphate from fructose-6-phosphate and ATP through domain closure that mimics adenylate kinase’s water-excluding motion, enabling efficient phosphoryl transfer. Conversely, the bisphosphatase activity hydrolyzes fructose-2,6-bisphosphate back to fructose-6-phosphate via a His-Pro-Pro motif nucleophilic attack that forms a transient phosphoenzyme intermediate, and reciprocal substrate inhibition ensures a rheostat-like steady-state function. Akt-dependent phosphorylation at Ser483 during PI3K/insulin signaling increases kinase activity to promote cardiac glycolysis after feeding, while PKA phosphorylation at Ser466/29 during β-adrenergic and cAMP elevation inhibits kinase and activates phosphatase, redirecting metabolism toward fat oxidation and providing ischemic protection, thus maintaining high basal fructose-2,6-bisphosphate for ATP homeostasis. Dysregulation of PFKFB2, including isoform switching or reduced expression, leads to diabetic cardiomyopathy with impaired contractility, glycolytic deficits in heart failure, and facilitates Warburg metabolic reprogramming in cancer.

Background

PFKFB2, the heart and muscle-predominant bifunctional enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase, precisely regulates glycolytic flux by controlling levels of fructose-2,6-bisphosphate, the most potent allosteric activator of phosphofructo-1-kinase, thereby accelerating glycolysis and inhibiting gluconeogenesis, a role especially vital in cardiac energy metabolism under stress, insulin, and adrenergic regulation. The enzyme forms a head-to-head homodimer, with each monomer containing an N-terminal regulatory segment, a central kinase domain with bi-lobed adenylate kinase-like architecture featuring a six-stranded β-sheet and α-helices, and an ATP/fructose-6-phosphate binding pocket with key catalytic residues that coordinate Mg²⁺-ATP. Its C-terminal phosphatase domain, homologous to histidine phosphatases, contains flexible loops that grant fructose-2,6-bisphosphate access for hydrolysis via a phosphohistidine relay, while the C-terminal tail harbors Ser466/483 sites for Akt and PKA phosphorylation. The kinase activity synthesizes fructose-2,6-bisphosphate from fructose-6-phosphate and ATP through domain closure that mimics adenylate kinase’s water-excluding motion, enabling efficient phosphoryl transfer. Conversely, the bisphosphatase activity hydrolyzes fructose-2,6-bisphosphate back to fructose-6-phosphate via a His-Pro-Pro motif nucleophilic attack that forms a transient phosphoenzyme intermediate, and reciprocal substrate inhibition ensures a rheostat-like steady-state function. Akt-dependent phosphorylation at Ser483 during PI3K/insulin signaling increases kinase activity to promote cardiac glycolysis after feeding, while PKA phosphorylation at Ser466/29 during β-adrenergic and cAMP elevation inhibits kinase and activates phosphatase, redirecting metabolism toward fat oxidation and providing ischemic protection, thus maintaining high basal fructose-2,6-bisphosphate for ATP homeostasis. Dysregulation of PFKFB2, including isoform switching or reduced expression, leads to diabetic cardiomyopathy with impaired contractility, glycolytic deficits in heart failure, and facilitates Warburg metabolic reprogramming in cancer.

References
https://pubmed.ncbi.nlm.nih.gov/27802586/ https://pubmed.ncbi.nlm.nih.gov/39763797/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%