Western blot analysis of extracts from various cell lines using Aldolase A (D73H4) Rabbit mAb.
Western blot analysis of SH-SY5Y Cell Extracts, untreated (-) or Enolase-1 knock-out (+) using Enolase-1 Antibody #3810 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from various cell lines and human and mouse brain using Enolase-2 Antibody.
Western blot analysis of extracts from various cell types using PDHK1 (C47H1) Rabbit mAb.
Confocal immunofluorescent analysis of LNCaP (high expression, left) and Hep G2 (low expression, right) cells using PFKFB2 (D7G5R) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Western blot analysis of extracts from various cell lines using PFKFB3 (D7H4Q) Rabbit mAb.
Western blot analysis of extracts from various cell lines using PGAM1 (D3J9T) Rabbit mAb.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from various cell types using Enolase-1 Antibody.
Immunoprecipitation of PFKFB2 from Hep G2 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or PFKFB2 (D7G5R) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using PFKFB2 (D7G5R) Rabbit mAb. An anti-rabbit IgG light chain antibody was used as the secondary antibody.
Western blot analysis of extracts from 293 cells, mock transfected (-) or transfected with a construct expressing DDK-tagged full-length human PFKFB3 (hPFKFB3-DDK; +), using PFKFB3 (D7H4Q) Rabbit mAb (upper), DYKDDDDK Tag (9A3) Mouse mAb #8146 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from LNCaP, Hep G2, and MCF7 cells using PFKFB2 (D7G5R) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
|Aldolase A (D73H4) Rabbit mAb 8060||20 µl||
||H Mk M||40||Rabbit IgG|
|Enolase-1 Antibody 3810||20 µl||
||H Mk M R||47||Rabbit|
|Enolase-2 Antibody 9536||20 µl||
|PDHK1 (C47H1) Rabbit mAb 3820||20 µl||
||H Mk M R||47||Rabbit IgG|
|PFKFB2 (D7G5R) Rabbit mAb 13045||20 µl||
||H Mk||55||Rabbit IgG|
|PFKFB3 (D7H4Q) Rabbit mAb 13123||20 µl||
||H Mk M R||60||Rabbit IgG|
|PGAM1 (D3J9T) Rabbit mAb 12098||20 µl||
||Hm H Mk M R||28||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
The Glycolysis II Antibody Sampler Kit provides an economical means to investigate select enzymes involved in glycolysis. The kit contains enough primary antibody to perform two western blot experiments per primary antibody.
Each antibody recognizes endogenous total levels of the specified target protein independent of its modified state.
Monoclonal antibodies are produced by immunizing animals with a synthetic peptides corresponding to residues surrounding Pro263 of human fructose bisphosphate aldolase A protein, human PDHK1 protein, residues surrounding Pro454 within the fructose-2,6-biphosphatase region of human PFKFB2 protein, residues surrounding Leu456 of human PFKFB3 protein, and the carboxy terminus of human PGAM1 protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of human enolase-1 and the carboxy terminus of human enolase-2 protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.
Glycolysis is the metabolic process by which glucose is converted to pyruvate in a sequence of enzymatic steps. Phosphofructokinase (PFK) catalyzes the phosphorylation of fructose-6-phosphate in glycolysis (1). The bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK-2/FBPase or PFKFB) catalyzes the synthesis and degradation of fructose 2,6-bisphosphate and regulates its steady-state level. Four different PFKFB isoforms (PFKFB1, PFKFB2, PFKFB3, and PFKFB4) have been identified (2). Aldolase (fructose bisphosphate aldolase) is a glycolytic enzyme that catalyzes the conversion of fructose 1, 6-bisphosphate to 3-phosphoglyceraldehyde (3). Phosphoglycerate mutase (PGAM1) catalyzes the conversion of 3-phosphoglycerate to 2-phosphoglycerate during glycolysis (4). Enolase is an important glycolytic enzyme involved in the interconversion of 2-phosphoglycerate to phosphoenolpyruvate. Mammalian enolase exists as three subunits: enolase-1 (α-enolase), enolase-2 (γ-enolase) and enolase-3 (β-enolase) that can form both homo- and heterodimers (5). Pyruvate dehydrogenase kinase (PDHK) phosphorylates PDH and inactivates it, whereas dephosphorylation of PDH is carried out by pyruvate dehydrogenase phosphatase to generate the active form (6).
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