Pyruvate Carboxylase Antibody

Western Blot: Pyruvate Carboxylase Antibody [NBP1-49536] - Analysis of Pyruvate Carboxylase in A. Human liver extracts and B. Mouse liver extracts

Immunocytochemistry/Immunofluorescence: Pyruvate Carboxylase Antibody [NBP1-49536] - Immunocytochemistry of Pyruvate Carboxylase in HeLa using NBP1-49536. Nuclei (Blue) are counterstained using Hoechst 33258.

Immunohistochemistry-Paraffin: Pyruvate Carboxylase Antibody [NBP1-49536] - IHC analysis of a formalin fixed and paraffin embedded tissue section of mouse kidney using rabbit anti-Pyruvate Carboxylase antibody at 1:200 dilution. The signal was developed using HRP conjugated anti-rabbit secondary antibody and DAB reagent. The nuclei were counterstained using hematoxylin. This Pyruvate Carboxylase antibody generated a diffused cytoplasmic staining in all the tubules and a subset of cells in glomerular cells.

Immunohistochemistry-Paraffin: Pyruvate Carboxylase Antibody [NBP1-49536] - IHC analysis of a formalin fixed and paraffin embedded tissue section of mouse adipose tissue using rabbit anti-Pyruvate Carboxylase antibody at 1:200 dilution. The signal was developed using HRP conjugated anti-rabbit secondary antibody and DAB reagent. The nuclei were counterstained using hematoxylin. This Pyruvate Carboxylase antibody generated a specific staining of the fat cells in the tested section.

Immunocytochemistry/Immunofluorescence: Pyruvate Carboxylase Antibody [NBP1-49536] - HeLa cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with anti-Pyruvate Carboxylase at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

• Reactivity: Hu, Mu, Rt, Rt, Po, Bv
• Applications: WB, ICC/IF, IHC, IHC-P
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

Pyruvate Carboxylase Antibody Summary

• Immunogen: A genomic peptide made to an internal region of the human Pyruvate Carboxylase protein (within residues 930-1050). [Swiss-Prot P11498]
• Localization: Mitochondrion matrix.
• Predicted Species: Rat (92%), Porcine (96%), Bovine (96%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: PC
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot
  • Immunocytochemistry/Immunofluorescence 1:50-1:100
  • Immunohistochemistry 1:200
  • Immunohistochemistry-Paraffin 1:200
• Application Notes: This Pyruvate Carboxylase antibody is useful for Immunohistochemistry on paraffin-embedded sections, Immunocytochemistry/Immunofluorescence and Western blot, where a band is seen at ~129 kDa. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. Use in Western blot reported in scientific literature (PMID: 29937374).
• Theoretical MW: 126 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  Human Liver Whole Tissue Lysate (Adult Whole Normal)
  Mouse Liver Whole Tissue Lysate (Adult Whole Normal)

Reactivity Notes

Human and mouse. Immunogen has 96% identity to bovine and porcine. Rat reactivity reported in scientific literature (PMID: 24333689)

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Tris-Glycine
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for Pyruvate Carboxylase Antibody

• EC 6.4.1
• EC 6.4.1.1
• PCBPyruvic carboxylase
• pyruvate carboxylase
• pyruvate carboxylase, mitochondrial

Background

Pyruvate Carboxylase (PC) catalyzes an important 2-step reaction - (i) ATP-dependent carboxylation of covalently attached biotin (ii) transfer of carboxyl group to pyruvate, and this interconversion is essential for replenishment of TCA/citric acid cycle intermediates (anaplerosis). By extension, PC participates in many pathways that depend on oxaloacetate (OAA) such as gluconeogenesis, glycogen synthesis, lipogenesis, glycerogenesis, glucose-dependent insulin secretion, and synthesis of amino acids as well as neurotransmitters. With highest expression levels in liver, adipose tissue, kidney, lactating mammary gland, pancreatic islets etc, PC localize to cellular mitochondrial matrix and its activity is highly regulated by its allosteric activator acetyl-CoA. Enhanced acetyl-CoA levels indicate a need for increased OAA synthesis to replenish TCA cycle intermediates or a diversion of pyruvate for glucose synthesis in gluconeogenic tissues. PC's deficiency causes a fatal multiorgan metabolic imbalance that predominantly manifests with lactic acidemia and neurological dysfunction at an early age. Three clinical forms of PC deficiency include an infantile or mild form (Type A), a severe neonatal form (Type B), and a benign form (Type C), all of which exhibit clinical or biochemical parallals of impaired anaplerosis.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Pyruvate Carboxylase Antibody (NBP1-49536)(9)

Publications using NBP1-49536

• Rattanapornsompong K, Ngamkham J, Chavalit T, Jitrapakdee S. Generation of Human Pyruvate Carboxylase Knockout Cell Lines Using Retrovirus Expressing Short Hairpin RNA and CRISPR-Cas9 as Models to Study Its Metabolic Role in Cancer Research Methods Mol. Biol. Dec 12 2019 [PMID: 30535704] (IHC, Human)
• Toledo M, Batista-Gonzalez A, Merheb E et al. Autophagy Regulates the Liver Clock and Glucose Metabolism by Degrading CRY1 Cell Metab. 2018 Aug 7 [PMID: 29937374] (Mouse)
• Toledo M, Batista-Gonzalez A, Merheb E et al. Autophagy Regulates the Liver Clock and Glucose Metabolism by Degrading CRY1 Cell Metab. 2018 Aug 7 [PMID: 29937374] (Mouse)
• Li X, Han G, Li X et al. Mitochondrial pyruvate carrier function determines cell stemness and metabolic reprogramming in cancer cells Oncotarget 2017 Jul 11 [PMID: 28624784] (WB, Mouse)
• Seale LA, Gilman CL, Hashimoto AC et al. Diet-induced obesity in the selenocysteine lyase knockout mouse Antioxid. Redox Signal. 2015 Jul 20 [PMID: 26192035]
• Tao H, Zhang Y, Zeng X et al. Niclosamide ethanolamine-induced mild mitochondrial uncoupling improves diabetic symptoms in mice. Nat. Med. 2014 Oct 05 [PMID: 25282357] (WB)
• Lee JH, Jung IR, Choi SE et al. Toxicity generated through inhibition of pyruvate carboxylase and carnitine palmitoyl transferase-1 is similar to high glucose/palmitate-induced glucolipotoxicity. Mol Cell Endocrinol 2013 Dec 11 [PMID: 24333689] (WB, Rat)
• Lee W, Reyes RC, Gottipati MK et al. Enhanced Ca2+-dependent glutamate release from astrocytes of the BACHD Huntington's disease mouse model. Neurobiol Dis 2013 Jun 10 [PMID: 23756199] (WB, Mouse)
• Vastag L, Koyuncu E, Grady SL et al. Divergent effects of human cytomegalovirus and herpes simplex virus-1 on cellular metabolism. PLoS Pathog. 2011 July. [PMID: 21779165] (WB, Human)

Bioinformatics

• Gene Symbol: PC
• Entrez:
  • 5091(Human)
• Uniprot:
  • P11498(Human)
  • Q05920(Mouse)