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12866
Glycolysis II Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Glycolysis II Antibody Sampler Kit #12866

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Citations (2)
Glycolysis II Antibody Sampler Kit: Image 1

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

Glycolysis II Antibody Sampler Kit: Image 2

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).

Glycolysis II Antibody Sampler Kit: Image 3

Western blot analysis of extracts from various cell lines and human and mouse brain using Enolase-2 Antibody.

Glycolysis II Antibody Sampler Kit: Image 4

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

Glycolysis II Antibody Sampler Kit: Image 5

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).

Glycolysis II Antibody Sampler Kit: Image 6

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

Glycolysis II Antibody Sampler Kit: Image 7

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

Glycolysis II Antibody Sampler Kit: 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.

Glycolysis II Antibody Sampler Kit: Image 9

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

Glycolysis II Antibody Sampler Kit: Image 10

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.

Glycolysis II Antibody Sampler Kit: Image 11

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).

Glycolysis II Antibody Sampler Kit: Image 12

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).

To Purchase # 12866T
Product # Size Price
12866T
1 Kit  (7 x 20 µl) $ 500

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Aldolase A (D73H4) Rabbit mAb 8060 20 µl
  • WB
H Mk M 40 Rabbit IgG
Enolase-1 Antibody 3810 20 µl
  • WB
  • IP
H Mk M R 47 Rabbit 
Enolase-2 Antibody 9536 20 µl
  • WB
H M 47 Rabbit 
PDHK1 (C47H1) Rabbit mAb 3820 20 µl
  • WB
  • IP
H Mk M R 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 Mk M R 60 Rabbit IgG
PGAM1 (D3J9T) Rabbit mAb 12098 20 µl
  • WB
Hm H Mk M R 28 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

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.

Specificity / Sensitivity

Each antibody recognizes endogenous total levels of the specified target protein independent of its modified state.

Source / Purification

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.

Background

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. Vander Heiden, M.G. et al. (2010) Science 329, 1492-9.
  5. Pancholi, V. (2001) Cell Mol Life Sci 58, 902-20.
  6. Wigfield, S.M. et al. (2008) Br J Cancer 98, 1975-84.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.