Pyruvate Carboxylase Antibody - BSA Free

Immunocytochemistry/Immunofluorescence: Pyruvate Carboxylase Antibody [NBP1-49536] - HeLa cells were fixed in 4% paraformaldehyde for 10 min and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti- (NBP1-49536) at 10 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

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

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.

Western Blot: Pyruvate Carboxylase Antibody [NBP1-49536] - Characterization of MPC1-/- in the RM-1 murine prostate cancer cells. Western blotting results of subcellular proteins from WT and MPC1-/- cells and corresponding densitometry histograms of ALT1 and Pyruvate Carboxylase. Cyto means cytoplasmic protein. Mito stands for mitochondrial protein. Protein alpha-Tublin was used as a loading control. ***P<0.001. All experiments were performed at least three times with consistent and repeatable results. Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.18199), licensed under a CC-BY license.

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.

Western Blot: Pyruvate Carboxylase Antibody [NBP1-49536] - HSV-1 replication is inhibited by reducing flux from glucose toward pyrimidine nucleotide synthesis. RNA interference knockdown of pyruvate carboxylase (marked by arrow) in MRC5 cells. Cells were transfected with non-targeting siRNAs (NT) or siRNAs targeting pyruvate carboxylase (PC) and harvested at indicated time points after transfection. Pyruvate carboxylase levels in the cells were detected by western blot using specific antibodies. Beta-actin was employed as a loading control. Divergent effects of human cytomegalovirus and herpes simplex virus-1 on cellular metabolism. PLoS Pathog (2011)

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

Pyruvate Carboxylase Antibody - BSA Free Summary

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

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:50-1:100
  • Immunohistochemistry 1:200
  • Immunohistochemistry-Paraffin 1:200
  • Knockdown Validated
  • Western Blot 1:1000. Use reported in scientific literature (PMID 29937374)
• Application Notes: In Western blot, 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.
• 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

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: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for Pyruvate Carboxylase Antibody - BSA Free

• 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)(17)

Publications using NBP1-49536

• Kershberg L, Banerjee A, Kaeser PS. Protein composition of axonal dopamine release sites in the striatum eLife 2022-12-29 [PMID: 36579890] (Western Blot, Immunofluorescence, Immunocytochemistry)
• Guo X, Jiang X, Chen K et al. The Role of Palmitoleic Acid in Regulating Hepatic Gluconeogenesis through SIRT3 in Obese Mice Nutrients 2022-04-01 [PMID: 35406095] (WB, Mouse)
• Schworer S, Pavlova NN, Cimino FV Et al. Fibroblast pyruvate carboxylase is required for collagen production in the tumour microenvironment Nature metabolism 2021-11-01 [PMID: 34764457] (WB, Human)
• Chung CY, Singh K, Kotiadis VN Et al. Constitutive activation of the PI3K-Akt-mTORC1 pathway sustains the G mtDNA mutation Nature communications 2021-11-04 [PMID: 34737295] (WB, Human)
• Igelmann S, Lessard F, Uchenunu O et al. A hydride transfer complex reprograms NAD metabolism and bypasses senescence Molecular cell 2021-09-16 [PMID: 34547241]
• Song X, Liu J, Kuang F et al. PDK4 dictates metabolic resistance to ferroptosis by suppressing pyruvate oxidation and fatty acid synthesis Cell reports 2021-02-23 [PMID: 33626342]
• Li X, Han G, Li X et al. Mitochondrial pyruvate carrier function determines cell stemness and metabolic reprogramming in cancer cells Oncotarget 2017-07-11 [PMID: 28624784] (WB, Mouse)
• Xin Cai, Charles P Ng, Olivia Jones, Tak Shun Fung, Keun Woo Ryu, Dayi Li, Craig B Thompson Lactate activates the mitochondrial electron transport chain independently of its metabolism. Molecular cell 2023-11-06 [PMID: 37879334]
• 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. 2019-12-12 [PMID: 30535704] (IF/IHC, Human)
• Toledo M, Batista-Gonzalez A, Merheb E et al. Autophagy Regulates the Liver Clock and Glucose Metabolism by Degrading CRY2 Cell Metab. 2018-08-07 [PMID: 29937374] (Mouse)
• Seale LA, Gilman CL, Hashimoto AC et al. Diet-induced obesity in the selenocysteine lyase knockout mouse Antioxid. Redox Signal. 2015-07-20 [PMID: 26192035]
• Chung C, Singh K, Kotiadis V et al. Maladaptive nutrient signalling sustains the m.3243A>G mtDNA mutation bioRxiv 2020-06-19 (WB, Human)
• Tao H, Zhang Y, Zeng X et al. Niclosamide ethanolamine-induced mild mitochondrial uncoupling improves diabetic symptoms in mice. Nat Med 2014-11-01 [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 2013-12-11 [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 2013-06-10 [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. 2011-07-01 [PMID: 21779165] (WB, Human)

Bioinformatics

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