View Featured Offers >>
67935
Electron Transport Chain (Complex II, III, IV) Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Electron Transport Chain (Complex II, III, IV) Antibody Sampler Kit #67935

Citations (0)
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.
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 SDHA (D6J9M) XP® Rabbit mAb.
Western blot analysis of extracts from various cell lines using COX10 (E6K4D) Rabbit mAb.
Western blot analysis of extracts from various cell lines using SDH5 (D1S8D) 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.
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.
Immunoprecipitation of SDHB protein from K-562 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is SDHB (E3H9Z) XP® Rabbit mAb. Western blot analysis was performed using SDHB (E3H9Z) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as the secondary antibody.
Western blot analysis of extracts from various cell lines and tissues using SDHB (E3H9Z) XP® Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). *The antibody detects a 120 kDa protein of unknown identity in some cell extracts.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using SDHB (E3H9Z) XP® 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.
Immunoprecipitation of SDHA from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or SDHA (D6J9M) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using SDHA (D6J9M) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing staining of the mitochondria, using COX IV (3E11) Rabbit mAb.
Western blot analysis of extracts from 293T cells, transfected with control siRNA (-) or SDHB siRNA (+), using SDHB (E3H9Z) XP® Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The loss of SDHB signal in extracts from SDHB siRNA-transfected cells confirms specificity of the antibody for SDHB.
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using SDHB (E3H9Z) XP® 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 colon carcinoma using SDHA (D6J9M) XP® 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).
Western blot analysis of extracts from 293T cells, mock transfected (lane 1) or transiently transfected with plasmid encoding Myc/DDK-tagged SDHB protein (lane 2), using SDHB (E3H9Z) XP® Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using SDHB (E3H9Z) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle using Cytochrome c (D18C7) Rabbit mAb.
Confocal immunofluorescent analysis of HeLa cells using SDHA (D6J9M) XP® Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded H1650 xenograft, using COX IV Rabbit mAb. Note specific staining of human cancer cells.
Immunohistochemical analysis of paraffin-embedded human ovarian clear cell carcinoma using SDHB (E3H9Z) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using SDHB (E3H9Z) XP® Rabbit mAb.
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).
Confocal immunofluorescent analysis of HCT 116 cells, either mock transfected (left, high-expressing) or transfected with siRNA directed against human SDHB (right, low-expressing) using SDHB (E3H9Z) XP® Rabbit mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
Immunohistochemical analysis of paraffin-embedded normal human heart using SDHB (E3H9Z) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded normal human kidney using SDHB (E3H9Z) XP® Rabbit mAb.
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.
Immunohistochemical analysis of paraffin-embedded Renca syngeneic tumor using SDHB (E3H9Z) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse colon using SDHB (E3H9Z) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse kidney using SDHB (E3H9Z) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse liver using SDHB (E3H9Z) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse ovary using SDHB (E3H9Z) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse stomach using SDHB (E3H9Z) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse testis using SDHB (E3H9Z) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse brain using SDHB (E3H9Z) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using SDHB (E3H9Z) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right). 
To Purchase # 67935
Cat. # Size Qty. Price
67935T
1 Kit

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
SDHA (D6J9M) XP® Rabbit mAb  11998 20 µl
  • WB
  • IP
  • IHC
  • IF
H M R Hm Mk 70 Rabbit IgG
SDHB (E3H9Z) XP® Rabbit mAb 92649 20 µl
  • WB
  • IP
  • IHC
  • IF
H M R Mk 26 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
SDH5 (D1S8D) Rabbit mAb 45849 20 µl
  • WB
  • IP
H M R Mk 15 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Rab Goat 

Product Description

The Electron Transport Chain (Complex II, III, IV) Antibody Sampler Kit provides an economical means of detecting select components involved in the electron transport chain (ETC) (Complex II, 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 II, III, IV) Antibody Sampler Kit detects endogenous levels of its target protein. SDHB (E3H9Z) XP® Rabbit mAb detects a 120 kDa protein of unknown identity in some cell extracts. 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 with synthetic peptides corresponding to residues surrounding Gly166 of human SDHA protein, Gly208 of human SDHB protein, Pro72 of human cytochrome c protein, Lys29 of human COX IV protein, Asp31 of human COX10 protein, Ala155 of human SDH5 protein, and near the carboxy terminus of 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

Succinate dehydrogenase (SDH), also known as Complex II or succinate:quinone oxidoreductase, is a key component of the citric acid cycle and the electron transport chain (ETC) (1). Specifically, it is involved in the oxidation of succinate (2). SDH consists of four subunits: SDHA, SDHB, SDHC, and SDHD (3). 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 (4). Cytochrome c is a well conserved electron transport protein and is part of the respiratory chain localized to mitochondrial intermembrane space (5). 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 (6). This event further triggers caspase-3 activation and eventually leads to apoptosis (7). 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 (8). 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 (9). Research studies show that the mRNAs of the mitochondrially encoded oxidative phosphorylation subunits, including COX1/MT-CO1, decline significantly during aging (10). Cytochrome c oxidase (COX) is a hetero-oligomeric enzyme consisting of 13 subunits localized to the inner mitochondrial membrane (11-13). 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 (14). COX10 is an assembly factor for cytochrome c oxidase (COX, also known as Complex IV) in the mitochondrial ETC (15,16). 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 (16). In addition, COX10 contributes to T cell quiescence exit and is critical for T cell activation (17). Succinate dehydrogenase subunit 5 (SDH5, SDHAF2) is a subunit of the succinate dehydrogenase (SDH) protein complex responsible for the oxidation of succinate during the citric acid cycle. Mitochondrial SDH5 associates with the catalytic subunit of succinate dehydrogenase and is required for adding FAD cofactor to the SDH catalytic subunit (18). Mutations in the corresponding SDHAF2 gene are associated with hereditary head and neck paragangliomas, an autosomal disorder characterized by the development of tumors with increased penetrance over time (18). Additional research studies show that SDH5 is involved in regulation of lung cancer metastasis mediated by the glycogen synthase kinase 3β and β-catenin signaling pathways (19).

  1. Oyedotun, K.S. and Lemire, B.D. (2004) J Biol Chem 279, 9424-31.
  2. Bourgeron, T. et al. (1995) Nat Genet 11, 144-9.
  3. Benchoua, A. et al. (2006) Mol Biol Cell 17, 1652-63.
  4. Maio, N. et al. (2017) Cell Metab 25, 945-953.e6.
  5. Schägger, H. (2002) Biochim Biophys Acta 1555, 154-9.
  6. Li, P. et al. (1997) Cell 91, 479-89.
  7. Liu, X. et al. (1996) Cell 86, 147-57.
  8. Nolfi-Donegan, D. et al. (2020) Redox Biol 37, 101674.
  9. Zong, S. et al. (2018) Cell Res 28, 1026-1034.
  10. Gomes, A.P. et al. (2013) Cell 155, 1624-38.
  11. Ostermeier, C. et al. (1996) Curr Opin Struct Biol 6, 460-6.
  12. Capaldi, R.A. et al. (1983) Biochim Biophys Acta 726, 135-48.
  13. Kadenbach, B. et al. (2000) Free Radic Biol Med 29, 211-21.
  14. Barrientos, A. et al. (2002) Gene 286, 53-63.
  15. Tarasenko, T.N. et al. (2017) Cell Metab 25, 1254-1268.e7.
  16. Zahalka, A.H. et al. (2017) Science 358, 321-326.
  17. Tan, H. et al. (2017) Immunity 46, 488-503.
  18. Hao, H.X. et al. (2009) Science 325, 1139-42.
  19. Liu, J. et al. (2013) J Biol Chem 288, 29965-73.

Pathways

Explore pathways related to this product.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not for Use in Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit our Trademark Information page.