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12866
Glycolysis II Antibody Sampler Kit

Glycolysis II Antibody Sampler Kit #12866

Western Blotting Image 1

Western blot analysis of extracts from various cell lines using Aldolase A (D73H4) Rabbit mAb.

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Western Blotting Image 2

Western blot analysis of extracts from various cell types using Enolase-1 Antibody.

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Western Blotting Image 3

Western blot analysis of extracts from various cell lines using Enolase-2 (D20H2) Rabbit mAb.

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Western Blotting Image 4

Western blot analysis of extracts from various cell types using PDHK1 (C47H1) Rabbit mAb.

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Western Blotting Image 5

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).

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Western Blotting Image 6

Western blot analysis of extracts from various cell lines using PFKFB3 (D7H4Q) Rabbit mAb.

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Western Blotting Image 7

Western blot analysis of extracts from various cell lines using PGAM1 (D3J9T) Rabbit mAb.

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Western Blotting Image 8

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.

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IP Image 9

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.

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Western Blotting Image 10

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).

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IF-IC Image 11

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).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Aldolase A (D73H4) Rabbit mAb 8060 20 µl
  • WB
H M Mk 40 Rabbit IgG
Enolase-1 Antibody 3810 20 µl
  • WB
  • IP
H M R Mk 47 Rabbit 
Enolase-2 (D20H2) Rabbit mAb 8171 20 µl
  • WB
  • IP
H M R Mk 47 Rabbit IgG
PDHK1 (C47H1) Rabbit mAb 3820 20 µl
  • WB
  • IP
H M R Mk 47 Rabbit IgG
PFKFB2 (D7G5R) Rabbit mAb 13045 20 µl
  • WB
  • IP
  • IF
H Mk 55 Rabbit IgG
PFKFB3 (D7H4Q) Rabbit mAb 13123 20 µl
  • WB
H M R Mk 60 Rabbit IgG
PGAM1 (D3J9T) Rabbit mAb 12098 20 µl
  • WB
H M R Hm Mk 28 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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, the carboxy terminus of human enolase-2 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 antibody is produced by immunizing animals with a synthetic peptide corresponding to the sequence of human enolase-1. 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).

  1. Mediavilla, D. et al. (2008) J Biochem 144, 235-44.
  2. Atsumi, T. et al. (2005) Diabetes 54, 3349-57.
  3. Castaldo, G. et al. (2000) Clin Chem 46, 901-6.
  4. Pancholi, V. (2001) Cell Mol Life Sci 58, 902-20.
  5. Vander Heiden, M.G. et al. (2010) Science 329, 1492-9.
  6. Wigfield, S.M. et al. (2008) Br J Cancer 98, 1975-84.
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

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