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72624
Gluconeogenesis Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Gluconeogenesis Antibody Sampler Kit #72624

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Gluconeogenesis Antibody Sampler Kit: Image 1
Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using GPI (E2Q8J) XP® Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 2
Immunohistochemical analysis of paraffin-embedded normal human colon using GPI (E2Q8J) XP® Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 3
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using GPI (E2Q8J) XP® Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 4
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using GPI (E2Q8J) XP® Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 5
Immunohistochemical analysis of paraffin-embedded normal human esophagus using GPI (E2Q8J) XP® Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 6
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin lymphoma using GPI (E2Q8J) XP® Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 7
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using GPI (E2Q8J) XP® Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 8
Immunohistochemical analysis of paraffin-embedded normal human salivary gland using GPI (E2Q8J) XP® Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 9
Western blot analysis of extracts from 293T cells, mock transfected (lane 1) or transiently transfected with plasmid encoding Myc/DDK-tagged GPI protein (lane 2), using GPI (E2Q8J) XP® Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Gluconeogenesis Antibody Sampler Kit: Image 10
Western blot analysis of extracts from various cell lines using GPI (E2Q8J) XP® Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Expression levels of GPI among cell lines are consistent with expectations based on publicly available bioinformatic databases, confirming specificity of the antibody for GPI.
Gluconeogenesis Antibody Sampler Kit: Image 11
Immunohistochemical analysis of paraffin-embedded TF-1 cell pellet (left, positive) or HuH-6 cell pellet (right, negative) using GPI (E2Q8J) XP® Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 12
Immunohistochemical analysis of paraffin-embedded normal human testis using GPI (E2Q8J) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Gluconeogenesis Antibody Sampler Kit: Image 13
Western blot analysis of extracts from various cell lines using PGAM1 (D3J9T) Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 14
Western blot analysis of extracts from various cell lines using PCK1 (D12F5) Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 15
Western blot analysis of extracts from mouse brain, rat brain, human brain, and mouse liver using Enolase-2 (E2H9X) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Gluconeogenesis Antibody Sampler Kit: Image 16
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).
Gluconeogenesis Antibody Sampler Kit: Image 17
Western blot analysis of extracts from MCF7 cells and various tissues using FBP1/FBPase 1 (D2T7F) Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 18
Western blot analysis of extracts from various cell lines using Pyruvate Carboxylase Antibody.
Gluconeogenesis Antibody Sampler Kit: Image 19
Western blot analysis of extracts from various cell lines using PGK1 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Gluconeogenesis Antibody Sampler Kit: Image 20
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.
Gluconeogenesis Antibody Sampler Kit: Image 21
Western blot analysis of extracts from various cell lines using PCK2 (D3E11) Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 22
Western blot analysis of extracts from 293 cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human PCK1 (hPCK1-Myc/DDK; +), using PCK1 (D12F5) Rabbit mAb (upper), DYKDDDDK Tag (9A3) Mouse mAb #8146 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Gluconeogenesis Antibody Sampler Kit: Image 23
Immunohistochemical analysis of paraffin-embedded human bladder adenocarcinoma using Enolase-2 (E2H9X) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Gluconeogenesis Antibody Sampler Kit: Image 24
Western blot analysis of extracts from various cell types using Enolase-1 Antibody.
Gluconeogenesis Antibody Sampler Kit: Image 25
Confocal immunofluorescent analysis of HeLa cells uisng PCK2 (D3E11) Rabbit mAb (green) showing colocalization with mitochondria that were labeled with MitoTracker® Red CMXRos (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Gluconeogenesis Antibody Sampler Kit: Image 26
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using Enolase-2 (E2H9X) XP® Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 27
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using Enolase-2 (E2H9X) XP® Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 28
Immunohistochemical analysis of paraffin-embedded human mucoepidermoid carcinoma of the larynx using Enolase-2 (E2H9X) XP® Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 29
Immunohistochemical analysis of paraffin-embedded mouse brain using Enolase-2 (E2H9X) XP® Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 30
Immunohistochemical analysis of paraffin-embedded mouse pancreas using Enolase-2 (E2H9X) XP® Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 31
Immunohistochemical analysis of paraffin-embedded TT cell pellet (left, positive) or Huh7 cell pellet (right, negative) using Enolase-2 (E2H9X) XP® Rabbit mAb.
Gluconeogenesis Antibody Sampler Kit: Image 32
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using Enolase-2 (E2H9X) XP® Rabbit mAb.
To Purchase # 72624
Cat. # Size Qty. Price
72624T
1 Kit  (6 x 20 microliters)
$ 650

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Enolase-1 Antibody 3810 20 µl
  • WB
  • IP
H M R Mk 47 Rabbit 
Enolase-2 (E2H9X) XP® Rabbit mAb 24330 20 µl
  • WB
  • IHC
H M R 47 Rabbit IgG
FBP1/FBPase 1 (D2T7F) Rabbit mAb 59172 20 µl
  • WB
H M R 39 Rabbit IgG
GPI (E2Q8J) XP® Rabbit mAb 94068 20 µl
  • WB
  • IHC
H Mk 60 Rabbit IgG
PCK1 (D12F5) Rabbit mAb 12940 20 µl
  • WB
H M R Mk 63 Rabbit IgG
PCK2 (D3E11) Rabbit mAb 8565 20 µl
  • WB
  • IP
  • IF
H Mk 71 Rabbit IgG
PGAM1 (D3J9T) Rabbit mAb 12098 20 µl
  • WB
H M R Hm Mk 28 Rabbit IgG
PGK1 Antibody 68540 20 µl
  • WB
H M R 43 Rabbit 
Pyruvate Carboxylase Antibody 66470 20 µl
  • WB
H M R Mk 130 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Gluconeogenesis Antibody Sampler Kit provides an economical means of detecting select components involved in the gluconeogenesis metabolism pathway. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Gluconeogenesis Antibody Sampler Kit detects endogenous levels of its target protein. Enolase-1 Antibody does not cross-react with enolase-2. Enolase-2 (E2H9X) XP® Rabbit mAb does not cross-react with human enolase-1 protein. PGAM1 (D3J9T) Rabbit mAb may cross-react with overexpressed PGAM2 protein.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues near the carboxy terminus of human enolase-2 protein, human PGAM1 protein, and human PCK1 protein, near the amino terminus of human PCK2 protein, and surrounding Val106 of human FBP1/FBPase 1 protein and Pro8 of human GPI protein.

Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Thr298 of human PGK1 protein, near the carboxy terminus of human pyruvate carboxylase protein, and corresponding to residues surrounding Ala76 of human enolase-1 protein. Antibodies are purified by protein A and peptide affinity chromatography. Pyruvate Carboxylase Antibody is purified by peptide affinity chromatography.




Background

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). Expression of the enolase isoforms differs in a tissue specific manner (1). Enolase-1 plays a key role in anaerobic metabolism under hypoxic conditions and may act as a cell surface plasminogen receptor during tissue invasion (2,3). Abnormal expression of enolase-1 is associated with tumor progression in some cases of breast and lung cancer (4-7). Alternatively, an enolase-1 splice variant (MBP-1) binds the c-myc promoter p2 and may function as a tumor suppressor. For this reason, enolase-1 is considered as a potential therapeutic target in the treatment of some forms of cancer (8). Research studies have shown elevated levels of neuron-specific enolase-2 in neuroblastoma (1) and small-cell lung cancer (9,10). Fructose-1,6-bisphosphatase 1 (FBP1 or FBPase 1), a rate-limiting enzyme in gluconeogenesis, catalyzes the conversion of fructose-1,6-bisphosphate to fructose-6-phosphate (11). Inhibition of FBP1 expression in basal-like breast cancer (BLBC) cells leads to metabolic reprogramming, including enhanced glycolysis, which leads to increased glucose uptake, biosynthesis of macromolecules, and activation of PKM2 (11). This metabolic reprogramming endows tumor cells with cancer stem cell (CSC)-like properties, thereby increasing their tumorigenicity (11). Depletion of FBP1 was also reported in more than 600 clear cell renal cell carcinoma (ccRCC) tumors, suggesting that FBP1 may inhibit ccRCC tumor progression (12). Glucose-6-phosphate isomerase (GPI) is a multi-functional protein belonging to the glucose phosphate isomerase family (13,14). As an intracellular metabolic enzyme, GPI plays a pivotal role in glycolysis and gluconeogenesis by catalyzing the interconversion of D-glucose-6-phosphate and D-fructose-6-phosphate (15). GPI is also secreted, where it functions as a cytokine (referred to as Autocrine Motility Factor, AMF), acting via the E3-ubiquitin-protein ligase AMFR/gp78 (16). In normal tissues, GPI/AMF has been shown to promote both immune cell maturation and neuronal cell survival (17,18). It is also secreted in abundance by some tumor cells (19), where it has been shown to promote tumor cell migration and metastasis (20,21). Phosphoenolpyruvate carboxykinase 1 (PCK1, PEPCK1, or PEPCK-C) is a cytosolic enzyme responsible for the conversion of oxaloacetate to phosphoenolpyruvate (22). PCK1 and PCK2 are involved in controlling the rate-limiting step of gluconeogenesis in the liver, which generates glucose from non-carbohydrate substrates, such as lactate and glycerol (23, 24). PCK2 (PEPCK2 or PEPCK-M) encodes an isoform of phosphoenolpyruvate carboxykinase (PEPCK) that is found in the mitochondria of renal and hepatic tissues (22). Phosphoglycerate mutase (PGAM1) catalyzes the conversion of 3-phosphoglycerate to 2-phosphoglycerate during glycolysis (25-29). Research studies have shown increased PGAM1 expression in cancer (25-28) and mental disease (29). PGK1 (phosphoglycerate kinase) is an essential enzyme in the glycolysis pathway (30). It catalyzes the reversible phospho-transfer reaction from 1,3-diphosphoglycerate to ADP to form ATP and 3-phosphoglycerate. The expression of PGK1 is upregulated in many cancer types and plays an important role in cancer cell proliferation and metastasis (31-34). Pyruvate carboxylase (PC) catalyzes the carboxylation of pyruvate to oxaloacetate to replenish TCA cycle intermediates. It is also critical in regulating gluconeogenesis in the liver (35).
  1. Pancholi, V. (2001) Cell Mol Life Sci 58, 902-20.
  2. Redlitz, A. et al. (1995) Eur J Biochem 227, 407-15.
  3. Jiang, B.H. et al. (1997) Cancer Res 57, 5328-35.
  4. Peebles, K.A. et al. (2003) Carcinogenesis 24, 651-7.
  5. Zhang, L. et al. (2000) J Surg Res 93, 108-19.
  6. Wu, W. et al. (2002) Clin Exp Metastasis 19, 319-26.
  7. Hennipman, A. et al. (1988) Tumour Biol 9, 241-8.
  8. Feo, S. et al. (2000) FEBS Lett 473, 47-52.
  9. Stern, P. et al. (2007) Tumour Biol 28, 84-92.
  10. O'Shea, P. et al. (1995) Ir J Med Sci 164, 31-6.
  11. Dong, C. et al. (2013) Cancer Cell 23, 316-31.
  12. Li, B. et al. (2014) Nature 513, 251-5.
  13. Haga, A. et al. (2000) Biochim Biophys Acta 1480, 235-44.
  14. Jeffery, C.J. et al. (2000) Biochemistry 39, 955-64.
  15. Kim, J.W. and Dang, C.V. (2005) Trends Biochem Sci 30, 142-50.
  16. Fairbank, M. et al. (2009) Mol Biosyst 5, 793-801.
  17. Gurney, M.E. et al. (1986) Science 234, 574-81.
  18. Gurney, M.E. et al. (1986) Science 234, 566-74.
  19. Lucarelli, G. et al. (2015) Medicine (Baltimore) 94, e2117.
  20. Liotta, L.A. et al. (1986) Proc Natl Acad Sci U S A 83, 3302-6.
  21. Funasaka, T. and Raz, A. (2007) Cancer Metastasis Rev 26, 725-35.
  22. Caton, P.W. et al. (2009) Life Sci 84, 738-44.
  23. Yoon, J.C. et al. (2001) Nature 413, 131-8.
  24. Fischer, S. et al. (2010) Biol Reprod 83, 859-65.
  25. Vander Heiden, M.G. et al. (2010) Science 329, 1492-9.
  26. Jacobowitz, D.M. et al. (2008) Microvasc Res 76, 89-93.
  27. Ren, F. et al. (2010) Mol Cancer 9, 81.
  28. Evans, M.J. et al. (2005) Nat Biotechnol 23, 1303-7.
  29. Martins-de-Souza, D. et al. (2009) BMC Psychiatry 9, 17.
  30. Beutler, E. (2007) Br J Haematol 136, 3-11.
  31. Wilson, R.B. et al. (2019) Pleura Peritoneum 4, 20190003.
  32. Yu, T. et al. (2017) Cancer Res 77, 5782-5794.
  33. Hu, H. et al. (2017) Hepatology 65, 515-528.
  34. Cao, H. et al. (2017) Cancer Chemother Pharmacol 79, 985-994.
  35. Cappel, D.A. et al. (2019) Cell Metab 29, 1291-1305.e8.

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