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42642
Electron Transport Chain (Complex I, III, IV) Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Electron Transport Chain (Complex I, III, IV) Antibody Sampler Kit #42642

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Simple Western analysis of lysates (1.0 mg/mL) from COS-7 cells using COX IV (3E11) Rabbit mAb #4850. The virtual lane view (left) shows the target bands (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess ​​​​​​​Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Immunoprecipitation of NDUFS1 protein from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is NDUFS1 (E4K3E) Rabbit mAb. Western blot analysis was performed using NDUFS1 (E4K3E) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human NDUFS1 (hNDUFS1-Myc/DDK; +), using NDUFS1 (E4K3E) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Confocal immunofluorescent analysis of A-673 cells using NDUFS1 (E4K3E) Rabbit mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
Western blot analysis of extracts from various cell lines using ACP/NDUFAB1 (E2V9B) Rabbit mAb.
Western blot analysis of extracts from various cell lines using Cytochrome c (D18C7) Rabbit mAb.
Western blot analysis of extracts from various cell lines using COX10 (E6K4D) Rabbit mAb.
Western blot analysis of extracts from HeLa, Jurkat and COS cell lines, using COX IV (3E11) Rabbit mAb.
Western blot analysis of extracts from various cell lines using COX1/MT-CO1 (E2I2R) Rabbit mAb.
Western blot analysis of extracts from various cell lines using COX1/MT-CO1 (E2I2R) Rabbit mAb.
Western blot analysis of extracts from various cell lines using NDUFS1 (E4K3E) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
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.
Western blot analysis of extracts from various cell lines using ACP/NDUFAB1 (E2V9B) Rabbit mAb.
Western blot analysis of extracts from various cell lines using UQCRFS1/RISP Antibody.
Western blot analysis of extracts from HeLa cells, untreated (-) or treated with Staurosporine #9953 (1 μM, 3 hr; +). Cells were fractionated into whole cell lysate (WCL), cytoplasm (Cyto), membrane (Mem), and cytoskeletal/nucleus (Nuc). Membrane fraction includes mitochondria.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing staining of the mitochondria, using COX IV (3E11) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Cytochrome c (D18C7) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using COX IV (3E11) Rabbit mAb in the presence of control peptide (left) or Cox IV Blocking Peptide #1034 (right).
Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle using Cytochrome c (D18C7) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded H1650 xenograft, using COX IV Rabbit mAb. Note specific staining of human cancer cells.
Confocal immunofluorescent analysis of HeLa cells labeled with COX IV (3E11) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Flow cytometric analysis of HeLa cells using COX IV (3E11) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
To Purchase # 42642
Cat. # Size Qty. Price
42642T
1 Kit

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
ACP/NDUFAB1 (E2V9B) Rabbit mAb 71814 20 µl
  • WB
H M Mk 9 Rabbit IgG
NDUFS1 (E4K3E) Rabbit mAb 70264 20 µl
  • WB
  • IP
  • IF
H M R 75 Rabbit IgG
UQCRFS1/RISP Antibody 95231 20 µl
  • WB
H M R 23 Rabbit 
Cytochrome c (D18C7) Rabbit mAb 11940 20 µl
  • WB
  • IHC
H M R Mk 14 Rabbit IgG
COX1/MT-CO1 (E2I2R) Rabbit mAb 55159 20 µl
  • WB
H M R Mk 32 Rabbit IgG
COX IV (3E11) Rabbit mAb 4850 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H R Mk Z B Pg 17 Rabbit IgG
COX10 (E6K4D) Rabbit mAb 24744 20 µl
  • WB
H Mk 49 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Rab Goat 

Product Description

The Electron Transport Chain (Complex I, III, IV) Antibody Sampler Kit provides an economical means of detecting select components involved in the electron transport chain (ETC) (Complex I, III, IV). The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Electron Transport Chain (Complex I, III, IV) Antibody Sampler Kit detects endogenous levels of its target protein. COX1/MT-CO1 (E2I2R) Rabbit mAb does not cross-react with COX2/MT-CO2 protein or COX3/MT-CO3 protein.

Source / Purification

Monoclonal antibodies are produced by immunizing animals synthetic peptides corresponding to residues surrounding Ile286 of human NDUFS1 protein, Pro72 of human cytochrome c protein, Lys29 of human COX IV protein, Asp31 of human COX10 protein, and near the carboxy terminus of human ACP/NDUFAB1 protein and human COX1/MT-CO1 protein.

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro86 of human UQCRFS1/RISP protein. Antibodies are purified by peptide affinity chromatography.

Background

Mitochondrial acyl carrier protein (ACP) is an essential component in fatty acid biosynthesis in mitochondria. It is also known as NADH-ubiquinone oxidoreductase subunit AB1 (NDUFAB1), a Complex I subunit in the electron transport chain (ETC). NDUFAB1 regulates energy and reactive oxygen species (ROS) metabolism in mitochondria by coordinating the assembly of ETC Complexes I, II, and III, and supercomplexes (1). NDUFS1 (NADH dehydrogenase Fe-S protein 1) is a nuclear-encoded structural subunit of NADH: ubiquinone oxidoreductase (Complex I) in the mitochondrial ETC (2). Mutations in NDUFS1 and other Complex I subunits leading to mitochondrial dysfunction are associated with a number of neurological disorders (3-6). Ubiquinol-cytochrome c reductase iron-sulfur subunit (UQCRFS1), also known as Rieske iron-sulfur protein (RISP), is a component of Complex III in the mitochondrial ETC. UQCRFS1/RISP and two other subunits, cytochrome b (MT-CYB) and cytochrome c1 (CYC1), are essential for the catalytic activity of Complex III (7). Cytochrome c is a well conserved electron transport protein and is part of the respiratory chain localized to mitochondrial intermembrane space (8). Upon apoptotic stimulation, cytochrome c released from mitochondria associates with procaspase-9 (47 kDa)/Apaf-1. This complex processes caspase-9 from inactive proenzyme to its active form (9). This event further triggers caspase-3 activation and eventually leads to apoptosis (10). The mitochondrial ETC comprises multiple protein complexes, including cytochrome c oxidase. Cytochrome c oxidase catalyzes the reduction of oxygen to water. This process is coupled with pumping protons from the mitochondrial matrix into mitochondrial intermembrane space, contributing to the proton gradient used for ATP synthesis (11). Cytochrome c oxidase consists of 3 mitochondrial DNA-encoded subunits (COX1/MT-CO1, COX2/MT-CO2, and COX3/MT-CO3) and multiple nuclear DNA-encoded subunits (12). Research studies show that the mRNAs of the mitochondrially encoded oxidative phosphorylation subunits, including COX1/MT-CO1, decline significantly during aging (13). Cytochrome c oxidase (COX, also known as Complex IV) is a hetero-oligomeric enzyme consisting of 13 subunits localized to the inner mitochondrial membrane (14-16). It is the terminal enzyme complex in the respiratory chain, catalyzing the reduction of molecular oxygen to water coupled to the translocation of protons across the mitochondrial inner membrane to drive ATP synthesis. The 3 largest subunits forming the catalytic core are encoded by mitochondrial DNA, while the other smaller subunits, including COX IV, are nuclear-encoded. Research studies have shown that deficiency in COX activity correlates with a number of human diseases (17). COX10 is an assembly factor for cytochrome c oxidase in the mitochondrial ETC (18,19). Studies show that, when the gene encoding the β2-adrenergic receptor (Adrb2) is deleted, increased oxidative phosphorylation in endothelial cells inhibits angiogenesis. Deletion of Cox10 prevents the metabolic switch to oxidative phosphorylation in endothelial cells deleted of Adrb2, causing angiogenesis and cancer progression (19). In addition, COX10 contributes to T cell quiescence exit and is critical for T cell activation (20).

  1. Hou, T. et al. (2019) Cell Res 29, 754-766.
  2. Duncan, A.M. et al. (1992) Cytogenet Cell Genet 60, 212-3.
  3. Petruzzella, V. et al. (2012) Adv Exp Med Biol 942, 371-84.
  4. Pagniez-Mammeri, H. et al. (2012) Mol Genet Metab 105, 163-72.
  5. Kashani, A. et al. (2014) Neurogenetics 15, 161-4.
  6. Zhu, Y. et al. (2015) J Hum Genet 60, 11-6.
  7. Maio, N. et al. (2017) Cell Metab 25, 945-953.e6.
  8. Schägger, H. (2002) Biochim Biophys Acta 1555, 154-9.
  9. Li, P. et al. (1997) Cell 91, 479-89.
  10. Liu, X. et al. (1996) Cell 86, 147-57.
  11. Nolfi-Donegan, D. et al. (2020) Redox Biol 37, 101674.
  12. Zong, S. et al. (2018) Cell Res 28, 1026-1034.
  13. Gomes, A.P. et al. (2013) Cell 155, 1624-38.
  14. Ostermeier, C. et al. (1996) Curr Opin Struct Biol 6, 460-6.
  15. Capaldi, R.A. et al. (1983) Biochim Biophys Acta 726, 135-48.
  16. Kadenbach, B. et al. (2000) Free Radic Biol Med 29, 211-21.
  17. Barrientos, A. et al. (2002) Gene 286, 53-63.
  18. Tarasenko, T.N. et al. (2017) Cell Metab 25, 1254-1268.e7.
  19. Zahalka, A.H. et al. (2017) Science 358, 321-326.
  20. Tan, H. et al. (2017) Immunity 46, 488-503.

Pathways

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