Antibodies to Mitochondria - Product Review 14

Cellular respiration is the process of oxidizing food molecules, like glucose, to carbon dioxide and water. The energy released is trapped in the form of ATP for use by all the energy-consuming activities of the cell.

The process occurs in two phases:

  • glycolysis, the breakdown of glucose to pyruvic acid
  • the complete oxidation of pyruvic acid to carbon dioxide and water

In eukaryotes, glycolysis occurs in the cytosol. The remaining processes take place in mitochondria.

Mitochondria are membrane-enclosed organelles distributed through the cytosol of most eukaryotic cells. Their main function is the conversion of the potential energy of food molecules into ATP. Mitochondria have:

  • an outer membrane that encloses the entire structure
  • an inner membrane that encloses a fluid-filled matrix ( between the two is the intermembrane spac, the inner membrane is elaborately folded with shelflike cristae projecting into the matrix)
  • a small number (some 5–10) circular molecules of DNA.

The matrix contains a complex mixture of soluble enzymes that catalyze the respiration of pyruvic acid and other small organic molecules.

At the centre of aerobic carbohydrate metabolism the pyruvate dehydrogenase complex (PDH) is localized in the matrix space of mitochondria where it catalyzes the irreversible oxidative decarboxylation of pyruvate entering the organelle to produce Acetyl-CoA, NADH and CO2. PDH complex deficiency is a relatively common cause of metabolic and neurodegenerative disorders. Enzyme defects are thought to contribute to late onset diabetes and the activity of the enzyme is altered by diet and exercise. Also PDH deficiency has been found in sepsis.

The inner membrane contains 5 complexes of integral membrane proteins:

  • NADH dehydrogenase (Complex I)
  • succinate dehydrogenase (Complex II)
  • cytochrome c reductase (Complex III; also known as the Cytochrome b-c1 complex)
  • cytochrome c oxidase (Complex IV)
  • ATP synthase (Complex V)

Complex I or NADH ubiquinone oxidoreductase is a large protein complex of 950,000 MW made up by 45-46 different subunits. The enzyme complex catalyses electron entry from NADH, as the name implies, via a flavin (FMN) and several non-heme iron centers. Complex I is sensitive to a wide range of inhibitors, many of which such as rotenone are pesticides or other common environmental toxins. Complex I dysfunction is a common cause of genetic OXPHOS defects. Altered functioning of this complex is also thought to contribute to several neurological disorders including Parkinsons disease and schizophrenia. Also there is evidence of complex I involvement in diabetes.

Complex II is also called succinate ubiquinone oxidoreductase or more commonly succinate dehydrogenase complex. This complex is composed of 4 subunits and contains a flavin (FAD), non-heme iron centers and a b-type cytochrome as prosthetic groups. It is both a component of the electron transport chain and an enzyme of the Krebs cycle. Complex II deficiencies are seen in OXPHOS genetic disease and found in a type of cancer called paraganglioma.

Complex III, or ubiquinol cytochrome c oxidoreductase, is a complex of 11 subunits and catalyses electron transfer from reduced ubiquinone or coenzyme Q 10 to cytochrome c. The complex contains 11 different subunits, one of which, cytochrome b, is encoded on mtDNA, the remainder are nuclear encoded. Genetic complex III deficiencies occur but are rare.

Complex IV, also called cytochrome c oxidoreductase or cytochrome c oxidase, is a complex of 13 different subunits. The complex contains 2 heme groups (a and a3) and 2 copper atoms as prosthetic groups. Genetic alterations of this enzyme complex are a common cause of OXPHOS diseases. The enzyme is altered in Alzheimers disease. Also there are reports of reduced amounts of this complex in cancer cells when these cells become hypoxic.

Complex V, also called F1F0ATPase and ATP Synthase, is responsible for ATP production in oxidative phosphorylation and can work in reverse as a proton pumping ATPase. The enzyme was thought to be localized exclusively to mitochondria. However, it has recently been identified also on the plasma membrane of several cell types including hepatocytes where it is the HDL receptor, on endothelial cells where it may act as the angiostatin receptor, and on the surface of cancer cells. The enzyme in mammals is composed of 17 subunits, 5 of which make up the easily detached F1. The remainder are components of 2 stalk domains and the proton pumping F0 part of the machinery. Two of the subunits of the F0 part are encoded on mtDNA while the other subunits are nuclear encoded. Mutations in the mt-encoded subunits of Complex V cause OXPHOS disease.

Antibody Tools for Detection of Complexes I-V and PDH

Acris Antibodies offers a wide range of products for the detection of mitochondrial complexes. All antibodies are thoroughly tested for immunoprecipitation, Western blotting and / or immunocytochemical applications. (See table and pictures below.)

Picture Gallery

Figure 1
Immunocytochemistry image of mitochondria using antibody MS110 (anti-NADH dehydrogenase subunit NDUFS3)

Immunocytochemistry image of mitochondria using antibody MS110 (anti-NADH dehydrogenase subunit NDUFS3)

Western blot using antibody MS104 (anti-NADH dehydrogenase subunit NDUFS4)

Western blot using antibody MS104 (anti-NADH dehydrogenase subunit NDUFS4)

Immunocytochemistry image of mitochondrion using antibody MS409 (anti-cytochrome c oxidase subunit 5a)

Immunocytochemistry image of mitochondrion using antibody MS409 (anti-cytochrome c oxidase subunit 5a)

Western blot using anti-F 1 F 0 ATPase -antibodies MS502, MS503, MS504, MS505

Western blot using anti-F 1 F 0 ATPase -antibodies MS502, MS503, MS504, MS505

Western blot using antibody MS507 (anti-F 1 F 0 ATPase Alpha/plant XR)

Western blot using antibody MS507 (anti-F 1 F 0 ATPase Alpha/plant XR)

Immunocytochemistry image of antibody MS507 (anti- F 1 F 0 ATPase Alpha/plant XR)

Immunocytochemistry image of antibody MS507 (anti- F 1 F 0 ATPase Alpha/plant XR)

Immunoprecipitation using anti-Pyruvate-dehydrogenase-antibody MSP01C

Immunoprecipitation using anti-Pyruvate-dehydrogenase-antibody MSP01C

Immunocytochemical staining with anti-Pyruvate-dehydrogenase-antibody MSP06: E2+E3 binding protein subunits (E2/E3bp)

Immunocytochemical staining with anti-Pyruvate-dehydrogenase-antibody MSP06: E2+E3 binding protein subunits (E2/E3bp)

Literature

Kristian T, Hopkins IB, McKenna MC, Fiskum G.Isolation of mitochondria with high respiratory control from primary cultures of neuros and astrocytes using nitrogen cavitation.J Neurosci Methods. 2005 Oct 24 (Epub ahead of print).

Capaldi RA, Murray J, Byrne L, Janes MS, Marushi MF.Immunological approaches to the characterization and diagnosis of mitochondrial disease.Mitochondrion. 2004 Sep; 4(5-6): 417-26.

Margineatu DH, Brown RM, Brown GK, Marcus AH, Capaldi RA.Heterogeneous distribution of pyruvate dehydrogenase in the matrix of mitochondria.Mitochondrion. 2002 Feb; 1(4): 327-38.

Chen YR, Chen CL, Zhang L, Green –Church KB, Zweier JL.Superoxide generation from mitochondrial NADH dehydrogenase induces self inactivation with specific protein radical formation.J Biol Chem. 2005 Sep 8; (Epub ahead of print).

Briere JJ, Favier J, Ghouzzi VE, Djouadi F, Benit P, Gimenez AP, Rustin P.Succinate dehydrogenase deficiency in human.Cell Mol Life Sci. 2005; 62(19-20):2317-24

Singh S, Khar A.Differential gene expression during apoptosis induced by a serum factor: Role of mitochondrialF(0)-F(1)ATP Synthase complex.Apoptosis. 2005 Oct. 3; (Epub ahead of print).

Yoshikawa S.Reaction mechanism and phospholipid structure of bovine heart cytochrome c oxidase.Biochem Soc Trans. 2005 Oct; 33(PT5): 934-7.

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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  

ATP5F1 antibody

Human MCF7; Host: Rabbit. Target Name: NOP56 . Sample Tissue: MCF7 . Antibody Dilution: 1.0ug/ml. ATP5F1 is supported by BioGPS gene expression data to be expressed in MCF7.; ATP5F1 antibody - middle region (ARP48188_P050) in Human MCF7 cells using Western Blot Rabbit Aff - Purified Bov, Can, Hu, Ms, Rt WB
50 µg / €350.00
  AVIVA Systems Biology

ATP5F1 antibody

Western  Blot:  ATP5F1 Antibody [NBP1-33465] – Western blot analysis of ATP5F1 on Molt-4 lysates using NBP1-33465 at a dilution of 1:1000. 30ug whole cell lysate loaded on 12% SDS PAGE gel. Rabbit IgG Hu ICC/IF, P, WB
0.1 ml / €420.00
  Novus Biologicals Inc.

ATPase subunit F6 antibody

ATPase subunit F6 Rabbit IgG Hu, Ms, Rt P, WB
0.1 mg / €400.00
  Novus Biologicals Inc.

ATP synthase subunit beta antibody

Human Brain; . Titration:2 ug/ml. Positive Control:Human brain stem cells; ATP5B antibody - C-terminal region (ARP48186_T100) in Human Brain cells using Immunohistochemistry Rabbit Aff - Purified African clawed frog, Bov, Chk, Hu, Ms, Rt, Ze P, WB
0.1 mg / €310.00
  AVIVA Systems Biology

ATP synthase subunit beta (Center) antibody

Formalin-fixed and paraffin-embedded human brain tissue reacted with ATP5B Antibody (Center), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. Rabbit Hu F, P, WB
0.4 ml / €370.00
  Acris Antibodies GmbH

ATP synthase subunit beta antibody

ATP synthase subunit beta Chicken Purified Bac, Plant Chloroplasts, Synechocystis C, WB
0.1 ml / €390.00
  AgriSera AB

ATP synthase subunit beta (WB Standard)

ATP synthase subunit beta Purified WB
0.25 ml / €270.00
  AgriSera AB

ATP synthase subunit beta antibody

ATP synthase subunit beta Rabbit Serum Arabidopsis, Bac, Glycine max, Hordeum vulgare, leafs, Lycopersicum esulentum, Maize, Oryza sp. (roots, Pl, pollen), Populus sp., Spinacia oleracea ICC/IF, WB
50 µl / €300.00
  AgriSera AB

ATP synthase subunit beta antibody

ATP synthase subunit beta Rabbit Serum Arabidopsis, Bac, Glycine max, Hordeum vulgare, leafs, Lycopersicum esulentum, Maize, Oryza sp. (roots, Pl, pollen), Populus sp., Spinacia oleracea WB
10 µl / €170.00
  AgriSera AB

ATP synthase subunit beta antibody

Human Muscle; ATP5B antibody - N-terminal region (ARP48185_T100) in Human Muscle cells using Immunohistochemistry Rabbit Aff - Purified Bov, Can, Hu, Ms, Por, Rb, Rt P, WB
0.1 mg / €310.00
  AVIVA Systems Biology

ATP synthase subunit beta antibody

Western Blot: E coli-derived fusion protein as test antigen. Affinity-purified IgY dilution: 1:2000, Goat anti-IgY-HRP dilution: 1:1000. Colorimetric method for signal development. ( anti-ATP5B ) Chicken Aff - Purified Hu, Ms, Rt WB
50 µg / €490.00
  Novus Biologicals Inc.

ATP synthase subunit beta antibody

Immunofluorescent: ATP Synthase beta Antibody (4.3E8.D10) [NB600-1171] - Figure 1 shows indirect immunofluorescence staining of rat neuronal/glial cell culture using NB600-1171. Mouse IgG1 4.3E8.D10 Purified Hu, Ms, Rt ICC/IF, IP, WB
0.1 mg / €400.00
  Novus Biologicals Inc.

ATP synthase subunit epsilon (N-term) antibody

ATP synthase subunit epsilon Rabbit Aff - Purified Bov, Hu, Ms, Por, Rt, Xen ICC/IF, WB
0.1 ml / €360.00
  Novus Biologicals Inc.

ATP synthase subunit epsilon (N-term) antibody

ATP synthase subunit epsilon Rabbit Aff - Purified Bov, Hu, Ms, Por, Rt, Xen ICC/IF, WB
25 µl / €160.00
  Novus Biologicals Inc.

Complex IV subunit III antibody

Human 721_B; WB Suggested Anti-COX3 Antibody Titration: 0.2-1 ug/ml. Positive Control: 721_B cell lysate; COX3 antibody - C-terminal region (ARP50251_P050) in Human 721_B cells using Western Blot Rabbit Aff - Purified Hu WB
50 µg / €350.00
  AVIVA Systems Biology

Complex IV subunit Va antibody

Immunohistochemistry-Paraffin: COX5A Antibody [NBP1-32550] - Immunohistochemical analysis of paraffin-embedded Cal27 xenograft, using COX5A(NBP1-32550) antibody at 1:500 dilution. Rabbit IgG Hu, Ms ICC/IF, P, WB
0.1 mg / €420.00
  Novus Biologicals Inc.

Complex IV subunit Vb antibody

Western Blot: COX5B Antibody [NBP1-31303] - Western blot analysis of COX5B Antibody on: A:Hep G2 15% SDS PAGE diluted at 1:1000 Rabbit IgG Hu E, P, WB
0.1 ml / €420.00
  Novus Biologicals Inc.

Complex IV subunit VIaH antibody

Western Blot: COX6a2 Antibody [NBP1-31112] - Western blot analysis of COX6a2 Antibody on: A:Raji 12% SDS PAGE diluted at 1:500 Rabbit IgG Hu Dot, P, WB
0.1 ml / €420.00
  Novus Biologicals Inc.

COX2b antibody

COX2b Rabbit Serum Chlamydomonas WB
0.1 ml / €390.00
  AgriSera AB

COX II antibody

COX II Chicken Purified Arabidopsis, Plantago, Silene WB
0.1 ml / €390.00
  AgriSera AB

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