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Product Information Revised: 18April2002 Monoclonal Antibodies Specific for Oxidative Phosphorylation Storage upon receipt: 26°C Protect from light All other products ≤ 20°C Desiccate Introduction Oxidative phosphorylation (OxPhos) occurs in the mitochondria and, in mammals, is catalyzed by five, membrane-bound protein complexes, namely NADH-ubiquinol oxidoreductase (Complex I), succinate-ubiquinol oxidoreductase (Complex II), ubiquinol-cytochrome c oxidoreductase (Complex III), cytochrome c oxidase (Complex IV) and ATP synthase (Complex V) (Figure 1). The complexes are composed of multiple subunits, some of which are encoded in the mitochondria while others are encoded in the nucleus. For example, mammalian cytochrome oxidase (COX) is composed of 13 subunits, three encoded by mitochondrial DNA (subunits, I, II and III) and ten encoded by nuclear DNA. Assembly of each complex involves a coordinated association of prosthetic groups (hemes, non-heme irons, flavins and copper atoms) with some polypeptides made in the mitochondrion and others made in the cytosol and then translocated to the organelle. This complicated process is poorly understood, but it requires various assembly factors, each of which is specific for a particular complex. Defects in assembly of one or more of these complexes contribute to several described mitochondrial diseases and possibly Alzheimers disease and Parkinsons disease.16 Molecular Probes offers a range of subunit-specific antibodies for the study of mitochondrial function and dysfunction (Tables 13 and Figure 1), including monoclonal antibodies specific for subunits of human COX (Complex IV) and for representative subunits of human Complexes I, II, III and V. These mouse monoclonal antibodies serve as important tools for investigating mitochondrial biogenesis and studying OxPhosrelated diseases. Patient cell lines can now be screened for deficiencies in each of the OxPhos complexes by simple Western blotting.7,8 When compared to control cell lines, this screen provides information about relative subunit expression levels and can be combined with native gel electrophoresis or sucrose gradient centrifugation to gather additional information regarding the assembly state of the OxPhos complex.9 Many of our antibodies against subunits of the OxPhos complexes may also be used for immunohistochemical analysis. Image analysis of the antibodys staining pattern can reveal the relative expression and localization of a subunit. This approach has been particularly useful for studying OxPhos subunit expression in diseased muscle fibers 10 and for screening Complex IVdeficient patients.11 For detection of these monoclonal antibodies, Molecular Probes offers antimouse IgG secondary antibodies labeled with a wide range of fluorophores, or with biotin, with enzymes such as horseradish peroxidase or alkaline phosphatase, with NANOGOLD® or Alexa Fluor® FluoroNanogold 1.4 nm gold clusters or with Captivate ferrofluid. For labeling mitochondrial targets in cells, the IgG1 antibodies in this group can be complexed with the reagents in our Zenon One Mouse IgG1 Labeling Kits. See our Web site (www.probes.com) for more information about these products. Figure 1. The mammalian oxidative phosphorylation system and specific monoclonal antibodies available from Molecular Probes. MP 06401 Monoclonal Antibodies Specific for Oxidative Phosphorylation Table 1. Specifications for Molecular Probes monoclonal antibodies specific for components of the oxidative phosphorylation system. Subunit Specificity Clone Isotype Suggested Conc. for Westerns (µg/mL) A-21344 39 kDa subunit 20C11 IgG1,k 0.5 NR A-21343 30 kDa subunit 3F9 IgG1,k 0.5 5 A-21360 20 kDa subunit 20E9 IgG1,k 0.5 NR 17 kDa subunit 21C11 IgG2b,k ND ND A-21342 15 kDa subunit 17G3 IgG1,k 0.1 5 A-21358 14 kDa subunit 21A6 IgG1,k 0.1 NR A-21357 8 kDa subunit 17C8 IgG1,k 1 5 70 kDa subunit 2E3 IgG1,k 0.1 0.5 30 kDa subunit 21A11 IgG2a,k 5 5 Core 1 subunit 16D10 IgG1,k 0.1 1 Core 2 subunit 13G12 IgG1,k 0.4 1 FeS subunit 5A5 IgG2b,k 0.5 2 A-21361 10 kDa subunit 1H9 IgG2a,k 0.2 ND A-6403 Subunit I 1D6 IgG2a,k 2 2 A-6404 Subunit II 12C4 IgG2a,k 2 5 A-6408 Subunit III § DA5 IgG2a,k 0.5 ND A-21348 Subunit IV 20E8 IgG2a,k 0.2 5 A-21347 Subunit IV 10G8 IgG2a,k 0.5 5 A-21363 Subunit Va 6E9 IgG2a,k 2 5 Subunit Vb 16H12 IgG2b,k 4 NR A-21364 Subunit VIa-H ** 4H2 IgG2a,k 5 ND A-21365 Subunit VIa-L ** 14A3 IgG1,k 5 NR A-21366 Subunit VIb 3F9 IgG1,k ND ND A-6401 Subunit VIc 3G5 IgG2b,k 2 5 A-21367 Subunit VIIa-H/L 6D7 IgG2a,k ND ND A-21368 Subunit VIIb 2G7 IgG1,k ND ND A-21350 a subunit 7H10 IgG2b,k 0.5 5 b subunit 3D5 IgG1,k 0.2 5 d subunit 7F9 IgG2b,k 5 0.5 A-21354 OSCP subunit 4C11 IgG1,k 0.1 1 A-21355 Inhibitor protein §§ 5E2 IgG1,k 0.3 5 Catalog Number A-21359 A-11142 A-21345 Oxidative Complex I II A-21362 A-11143 A-21346 A-21349 III IV (COX) A-21351 A-21353 V (ATPase) Suggested Conc. for IHC (µg/mL) Species Specificity * Hum Bov Rat Mou + + + + + + + + + + + + + + + + + + + + ± ± + + + + + + + + + + + + + + + + + + + ND ND ND ND ND ND ND ± ± + + ND ND ND ND + + + + ± + ND ND ND ND ND + ± + ± ND ND ND + + + + + + + + + + + + + + ± + ± + + + + ND ND ND ND ND + ND ND ND ND ND ND ND + ND ND ND ND ND ND ND ND + + ND * Species specificity: Hum, human; Bov, bovine; Mou, mouse; (+), strong reactivity; (±), weak reactivity; -(), no reactivity. When these monoclonal antibodies are used in immunohistochemistry applications, formaldehyde-fixed target cells may require antigen-retrieval methods before the target cells are permeabilized and blocked. These monoclonal antibodies yield the strongest and most consistent signals when used for indirect immunocytochemistry.§ This antibody was raised against the yeast subunit, but strongly crossreacts with human COX subunit III. ** The designations VIa-H and VIa-L refer to COX subunit VIa from bovine heart and liver, respectively. This antibody recognizes COX subunit VIIa isotypes from both heart and ilver. OSCP = Oligomyosin SensitivityConferring Protein. §§ Although not considered a member of Complex V, the ATPase inhibitor protein does associate with this OxPhos subsystem. ND = not determined. NR = not recommended. 2 Monoclonal Antibodies Specific for Oxidative Phosphorylation Table 2. Molecular characteristics of those components of the oxidative phosphorylation system for which Molecular Probes offers antibodies. Complex I II III IV V Complex Name NADH-ubiquinone oxidoreductase Succinate-ubiquinone oxidoreductase Ubiquinonecytochrome c Cytochrome c oxidase (COX) ATP synthase (F1F0) Subunit Name Molecular Weight * Alternate Name Protein Number Gene Number Catalog Number 39 kDa subunit 42.5 alpha subcomplex, 9 Q16795 NDUFA9 A-21344 30 kDa subunit 30.2 ironsulfur protein, 3 O75489 NDUFS3 A-21343 20 kDa subunit 26.6 ironsulfur protein, 7 O75251 NDUFS7 A-21360 17 kDa subunit 15.5 beta subcomplex, 6 O95139 NDUFB6 A-21359 15 kDa subunit 12.5 ironsulfur protein, 5 O43920 NDUFS5 A-21342 14 kDa subunit 15.1 alpha subcomplex, 6 P56556 NDUFA6 A-21358 8 kDa subunit 10.9 alpha subcomplex, 2 O43678 NDUFA2 A-21357 70 kDa subunit 72.7 flavoprotein P31040 SDHA A-11142 30 kDa subunit 31.6 ironsulfur protein P21912 SDHB A-21345 Core 1 subunit 51.6 none P31930 UQCRC1 A-21362 Core 2 subunit 48.5 none P22695 UQCRC2 A-11143 FeS subunit 29.6 Rieske ironsulfur protein P47985 UQCRFS1 A-21346 10 kDa subunit 9.6 § subunit VII P13271 § UCRQ A-21361 Subunit I 57.0 COX I P00395 MTCO1 A-6403 Subunit II 25.6 COX II P00403 MTCO2 A-6404 Subunit III 29.9 COX III P00414 MTCO3 A-6408 Subunit IV 19.6 COX IV P13073 COX4 A-21348 Subunit Va 16.8 COX Va P20674 COX5A A-21363 Subunit Vb 13.7 COX Vb P10606 COX5B A-21249 Subunit VIa-H ** 10.8 § COX VIa P07471 § COX6A2 A-21364 Subunit VIa-L ** 9.5 § COX VIa P13182 § COX6A1 A-21365 Subunit VIb 10.0 § COX VIb P00429 § COX6B A-21366 Subunit VIc 8.8 COX VIc P09669 COX6C A-6401 Subunit VIIa-H/L ** 9.4 COX VIIa P14406 COX7A2 A-21367 Subunit VIIb 9.2 COX VIIb P24311 COX7B A-21368 a subunit 59.8 F1 complex, alpha subunit P25705 ATP5A1 A-21350 b subunit 56.6 F1 complex, beta subunit P06576 ATP5B A-21351 OSCP subunit 23.2 F1 complex, O subunit P48047 ATP5O A-21354 d subunit 18.4 none O75947 ATP5H A-21353 Inhibitor protein (IP) 12.3 § IF1 O01096 ATPI A-21355 * Molecular weight, in kilodaltons, based upon the human protein sequence unless otherwise noted; many of these proteins show aberrant molecular weights when analyzed by SDS polyacrylamide gel electrophoresis. Protein reference numbers are from SWISS-PROT, an annotated protein sequence database (http://www.ebi.ac.uk/swissprot/) and refer to the human protein sequence, unless otherwise noted. Gene reference numbers are from Genbank, which can be accessed through Entrez (http://www3.ncbi.nlm.nih.gov/Entrez/). § Protein molecular weight and protein number are for the bovine sequence. ** H = heart-specific isotype; L = liver-specific isotype; H/L = recognizes COX subunit VIIa isotypes from both heart and liver. OSCP = Oligomycin sensitivityconferring protein. 3 Monoclonal Antibodies Specific for Oxidative Phosphorylation Monoclonal Antibodies Specific for OxPhos Complex IV (Cytochrome Oxidase) Table 3. Alexa Fluor labeled anti-Cox, subunit I antibodies.* Catalog Number To facilitate the study of cytochrome oxidase (COX) structure and mitochondrial biogenesis, Molecular Probes offers subunitspecific mouse antiOxPhos Complex IV monoclonal antibodies (Tables 1 and 2). COX catalyzes the transfer of electrons from reduced cytochrome c to molecular oxygen, with a concomitant translocation of protons across the mitochondrial inner membrane.12,13 This mitochondrial membranebound enzyme is composed of subunits that are encoded in both the mitochondria (COX subunits I, II and III) and the nucleus (all others). The binding specificity exhibited by our antiOxPhos Complex IV monoclonal antibody preparations allows researchers to investigate the regulation, assembly and orientation of COX subunits from a variety of organisms,1418 and the antibodies have proven valuable for analyzing human mitochondrial myopathies and related disorders.7,10,19,20 Alexa Fluor 488 and Alexa Fluor 594 conjugates of antiCOX subunit I are also available for direct staining of mitochondria (Table 3). Fluorophore Abs Em A-21296 Alexa Fluor 488 495 519 A-21297 Alexa Fluor 594 590 617 * For best results in immunohistochemistry, we have found that an antigenretrieval procedure is required. Approximate absorption (Abs) and fluorescence emission (Em) maxima, in nm, for conjugates. Materials Contents Molecular Probes unlabeled monoclonal antibodies specific for subunits of human OxPhos complexes are supplied in unit sizes of 50 µg or 100 µg. The antiyeast COX subunit III monoclonal antibody (A-6408), which crossreacts with the corresponding human subunit, is supplied in a unit size of 250 µg. The antihuman mitochondrial porin monoclonal antibody (A-21317) is supplied in a unit size of 100 µg. To prepare stock solutions of the lyophilized antibodies, reconstitute in 0.51 mL of phosphate-buffered saline (PBS), pH 7.4, containing 1% BSA. The Alexa Fluor dyelabeled antiCOX subunit I antibodies are supplied in unit sizes of 100 µL as 1 mg/mL solutions in 0.1 M sodium phosphate, 0.1 M NaCl, pH 7.5, 5 mM sodium azide. The degree of labeling for each conjugate is typically 28 molecules per IgG molecule; the exact degree of labeling is indicated on the product label. At the time of preparation, the products are certified to be free of unconjugated dyes. Monoclonal Antibodies Specific for Complexes I, II, III and V Molecular Probes offers a large number of monoclonal antibodies specific for individual subunits of Complexes I, II, III and V, as well as for the Complex V inhibitor protein (Tables 1 and 2). When these monoclonal antibodies are used in combination with the set of antibodies to cytochrome oxidase (Complex IV), the relative levels of all OxPhos enzyme complexes in normal and diseased tissues can be evaluated. The antiOxPhos Complex V subunit a, mouse monoclonal 7H10 (antiF1F0-ATPase subunit a, A-21350) and a polyclonal antibody comparable to antiOxPhos Complex V subunit b, mouse monoclonal 3D5 (antiF1F0-ATPase subunit b, A-21351) have also been shown to mimic angiostatin, a potent inhibitor of angiogenesis.21 Angiostatin protein (A-23375), a recombinant form of natural angiostatin, targets the F1F0-ATP synthase and inhibits cell-surface ATP metabolism of endothelial cells, thereby blocking cell migration and proliferation that is essential for angiogenesis. This research demonstrated that these anti-ATPase antibodies had similar inhibitory effects, implying that they also compromised ATP metabolism and may function as angiostatin analogs. Storage Upon receipt, store the lyophilized antibodies desiccated at ≤20°C. When properly stored, these products are stable for at least one year. Store solutions of the antibodies for up to two weeks at 26°C with the addition of 2 mM sodium azide. For longer storage, divide solutions into single-use aliquots and freeze at ≤20°C. AVOID REPEATED FREEZING AND THAWING. Upon receipt of the Alexa Fluor dyelabeled antiCOX subunit I antibodies, store at 26°C. When properly stored, these products are stable for at least six months. For longer storage, divide solutions into single-use aliquots and freeze at ≤20°C. AVOID REPEATED FREEZING AND THAWING, AND PROTECT FROM LIGHT. Related Products: Mitochondrial Porin and Mitochondrial Protein Extracts Mitochondrial porin is an outer-membrane protein that forms regulated channels (referred to as Voltage Dependent Anionic Channels, or VDACs) between the cytosol and the mitochondrial inter-membrane space.22 This abundant transmembrane protein forms a small pore (~3 nm) in the outer membrane, allowing molecules less than ~10 kDa to pass.23 Due to its abundance, porin is often used as a reference standard in Western blots when assaying for other mitochondrial proteins 9,24 and serves as an effective organelle marker for immunohistochemistry.25 A monoclonal antibody against human porin is available (A-21317). For researchers seeking a source of mitochondrial protein standards, Molecular Probes offers human heart mitochondrial proteins for SDS-polyacrylamide gel electrophoresis (M-22430) and for 2-D gel electrophoresis (M-22431). These products are complete mitochondrial lysates that have tested negative for hepatitis B and C, as well as HIV 1 and 2, in serology tests. These mitochondrial protein extracts are useful for comparing new mitochondrial protein preparations in either 1-D or 2-D gels and for testing mitochondrial antibodies. Applications The OxPhos antibodies bind monospecifically to the corresponding protein complex of the human oxidative phosphorylation system (Table 1 and Figure 1). These antibodies are versatile and can be used in a variety of assay formats, including solid-phase binding of native proteins, Western blots of denatured proteins and immunohistochemistry of fixed tissues. Several of the antibodies have been used in studies of human mitochondrial myopathies 7,10,26 and may prove to be useful tools not only for experimental cell biology but also for research on human disease states. Because staining protocols vary with application, the appropriate dilution of an antibody should be determined empirically (see Table 1). For the antihuman mitochondrial porin antibody, a final concentration of 0.11 µg/mL is suggested for both West4 Monoclonal Antibodies Specific for Oxidative Phosphorylation proteins involved in the oxidative phosphorylation system are integral membrane proteins, which contain a large number of hydrophobic amino acids that are likely to cause the protein to run aberrantly, with respect to their molecular weight, in SDS polyacrylamide gel electrophoresis. ern blots and immunohistochemical applications. For the fluorophore-labeled antibodies, a final concentration of 110 µg/mL should be satisfactory for most immunohistochemical applications.27 It is a good practice to centrifuge the Alexa Fluor dye labeled protein conjugate solutions briefly in a micro-centrifuge before use; only the supernatants should then be added to the experiment. This step will eliminate any protein aggregates that may have formed during storage, thereby reducing nonspecific background staining. Antigen-Retrieval In immunohistochemistry applications, formaldehyde-fixed target cells may require antigen-retrieval methods before the target cells are permeabilized and blocked (see Table 1). Specifications 1. Perform antigen retrieval by submersing the slide in 50 mM Tris, pH 8.0, in a staining jar and then placing this jar in a boiling-water bath for 30 minutes. Add boiling chips to the boilingwater bath to prevent splattering. The OxPhos monoclonal antibodies were prepared against the appropriate OxPhos complexes, isolated from bovine heart, bovine liver or human heart. The following stringent selection criteria were applied during the development of these mAbs: 1) ability of the antibody to detect native protein by particle-concentration fluorescence immunoassay (PCFIA) and enzyme-linked immunoabsorbent assay (ELISA), 2) monospecificity of the antibody for denatured protein in Western blots of whole-cell extracts and isolated mitochondria and 3) specific mitochondrial subcellular localization of the antibody in fixed cultured human cells. The purity and yield of each preparation was assessed with SDS-polyacrylamide gel electrophoresis and quantitative immunoassay specific for mouse IgG of the appropriate isotype. The subunit specificity and immunoglobin isotype are shown in Table 1. Note: Most of the 2. Cool the slide to room temperature. 3. Wash the specimen for 10 minutes in PBS plus 0.1% Triton® X-100. 4. Wash the specimen twice for 5 minutes each in PBS. 5. Block the specimen for 30 minutes in BlockAid blocking solution (B-10710) or in another suitable blocking formulation. References 1. Biochim Biophys Acta 1366, 199 (1998); 2. Biochim Biophys Acta 1366, 211 (1998); 3. Curr Opin Cardiol 13, 190 (1998); 4. Ann Neurol 44, S99 (1998); 5. J Neural Transm 105, 855 (1998); 6. Seminars in Liver Disease 18, 237 (1998); 7. Biochim Biophys Acta 1362, 145 (1997); 8. J Biol Chem 276, 8892 (2001); 9. J Biol Chem 276, 16296 (2001); 10. Biochim Biophys Acta 1315, 199 (1996); 11. Brain 123, 591 (2000); 12. Science 283, 1488 (1999); 13. Annu Rev Biochem 59, 569 (1990); 14. Biochemistry 30, 3674 (1991); 15. Biochim Biophys Acta 1225, 95 (1993); 16. Methods Enzymol 260, 117 (1995); 17. J Biol Chem 268, 18754 (1993); 18. J Biol Chem 266, 7688 (1991); 19. Pediatr Res 28, 529 (1990); 20. Hum Mol Genet 6, 935 (1997); 21. Proc Natl Acad Sci USA 98, 6656 (2001); 22. Biochim Biophys Acta 894, 109 (1987); 23. J Biol Chem 273, 24406 (1998); 24. Biochim Biophys Acta 1455, 35 (1999); 25. Mol Biol Cell 9, 917 (1998); 26. Pediatric Res 28, 529 (1990); 27. Short Protocols in Molecular Biology, 2nd Edition, F.M. Ausubel et al., Eds., John Wiley and Sons (1992) pp. 14-2414-30. Product List Cat # A-21358 A-21342 A-21359 A-21360 A-21343 A-21344 A-21357 A-21345 A-11142 A-21361 A-21362 A-11143 A-21346 A-21296 A-21296 A-6403 A-6404 A-6408 Current prices may be obtained from our Web site or from our Customer Service Department. Product Name Unit Size anti-OxPhos Complex I 14 kDa subunit, mouse IgG1, monoclonal 21A6 *human reactivity* .................................................... 100 µg anti-OxPhos Complex I 15 kDa subunit, mouse IgG, monoclonal 17G3 *human reactivity* ..................................................... 100 µg anti-OxPhos Complex I 17 kDa subunit, mouse IgG2b, monoclonal 21C11 *human reactivity* ................................................ 100 µg anti-OxPhos Complex I 20 kDa subunit, mouse IgG1, monoclonal 20E9 *human reactivity* .................................................... 100 µg anti-OxPhos Complex I 30 kDa subunit, mouse IgG1, monoclonal 3F9 *human reactivity* ...................................................... 100 µg anti-OxPhos Complex I 39 kDa subunit, mouse IgG1, monoclonal 20C11 *human reactivity* .................................................. 100 µg anti-OxPhos Complex I 8 kDa subunit, mouse IgG1, monoclonal 17C8 *human reactivity* ...................................................... 100 µg anti-OxPhos Complex II 30 kDa subunit, mouse IgG2a, monoclonal 21A11 *human reactivity* ................................................ 100 µg anti-OxPhos Complex II 70 kDa subunit, mouse IgG1, monoclonal 2E3 *human reactivity* ..................................................... 50 µg anti-OxPhos Complex III 10 kDa subunit, mouse IgG2a, monoclonal 1H9 *human reactivity* .................................................. 100 µg anti-OxPhos Complex III core 1 subunit, mouse IgG1, monoclonal 16D10 *human reactivity* ................................................. 100 µg anti-OxPhos Complex III core 2 subunit, mouse IgG1, monoclonal 13G12 *human reactivity* ................................................. 100 µg anti-OxPhos Complex III subunit FeS, mouse IgG2b, monoclonal 5A5 *human reactivity* ....................................................... 100 µg anti-OxPhos Complex IV subunit I, mouse IgG2a, monoclonal 1D6, Alexa Fluor® 488 conjugate (anti-cytochrome oxidase subunit I, Alexa Fluor ® 488 conjugate) *1 mg/mL* *human reactivity* ............................................. 100 µg anti-OxPhos Complex IV subunit I, mouse IgG2a, monoclonal 1D6, Alexa Fluor® 594 conjugate (anti-cytochrome oxidase subunit I, Alexa Fluor ® 594 conjugate) *1 mg/mL* *human reactivity* ............................................. 100 µg anti-OxPhos Complex IV subunit I, mouse IgG2a, monoclonal 1D6 (anti-cytochrome oxidase subunit I) *human reactivity* ... 100 µg anti-OxPhos Complex IV subunit II, mouse IgG2a, monoclonal 12C4 (anti-cytochrome oxidase subunit II) *human reactivity* ..................................................................................................................................................................... 100 µg anti-OxPhos Complex IV subunit III (yeast), mouse IgG2a, monoclonal DA5 (anti-cytochrome oxidase subunit III (yeast)) ..... 250 µg 5 Monoclonal Antibodies Specific for Oxidative Phosphorylation A-21347 A-21348 A-21363 A-21349 A-21364 A-21365 A-21366 A-6401 A-21367 A-21368 A-21355 A-21350 A-21351 A-21353 A-21354 A-21317 anti-OxPhos Complex IV subunit IV, mouse IgG2a, monoclonal 10G8 (anti-cytochrome oxidase subunit IV) *human reactivity* .................................................................................................................................................................... anti-OxPhos Complex IV subunit IV, mouse IgG2a, monoclonal 20E8 (anti-cytochrome oxidase subunit IV) *human reactivity* ..................................................................................................................................................................... anti-OxPhos Complex IV subunit Va, mouse IgG2a, monoclonal 6E9 (anti-cytochrome oxidase subunit Va) *human reactivity* ..................................................................................................................................................................... anti-OxPhos Complex IV subunit Vb, mouse IgG2b, monoclonal 16H12 (anti-cytochrome oxidase subunit Vb) *human reactivity* ..................................................................................................................................................................... anti-OxPhos Complex IV subunit VIa-H, mouse IgG2a, monoclonal 4H2 (anti-cytochrome oxidase subunit VIa-H) *human reactivity* ..................................................................................................................................................................... anti-OxPhos Complex IV subunit VIa-L, mouse IgG1, monoclonal 14A3 (anti-cytochrome oxidase subunit VIa-L) *human reactivity* .................................................................................................................................................................... anti-OxPhos Complex IV subunit VIb, mouse IgG1, monoclonal 3F9 (anti-cytochrome oxidase subunit VIb) *human reactivity* ..................................................................................................................................................................... anti-OxPhos Complex IV subunit VIc, mouse IgG2b, monoclonal 3G5 (anti-cytochrome oxidase subunit VIc) *human reactivity* ..................................................................................................................................................................... anti-OxPhos Complex IV subunit VIIa-H/L, mouse IgG2a, monoclonal 6D7 (anti-cytochrome oxidase subunit VIIa-H/L) *human reactivity* .................................................................................................................................................................... anti-OxPhos Complex IV subunit VIIb, mouse IgG1, monoclonal 2G7 (anti-cytochrome oxidase subunit VIIb) *human reactivity* ..................................................................................................................................................................... anti-OxPhos Complex V inhibitor protein, mouse IgG1, monoclonal 5E2 (anti-ATP synthase IP; anti-F1F0-ATPase IP) *human reactivity* ..................................................................................................................................................................... anti-OxPhos Complex V subunit a, mouse IgG2b, monoclonal 7H10 (anti-ATP synthase subunit a; anti-F1F0-ATPase subunit a) *human reactivity* .................................................................................................................................................... anti-OxPhos Complex V subunit b, mouse IgG1, monoclonal 3D5 (anti-ATP synthase subunit b; anti-F1F0-ATPase subunit b) *human reactivity* .................................................................................................................................................... anti-OxPhos Complex V subunit d, mouse IgG2b, monoclonal 7F9 (anti-ATP synthase subunit d; anti-F1F0-ATPase subunit d) *human reactivity* .................................................................................................................................................... anti-OxPhos Complex V subunit OSCP, mouse IgG1, monoclonal 4C11 (anti-ATP synthase subunit OSCP; anti-F1F0-ATPase subunit OSCP) *human reactivity* ............................................................................................................... anti-porin (human mitochondrial), mouse IgG2b, monoclonal 20B12 .......................................................................................... 100 µg 100 µg 100 µg 100 µg 100 µg 100 µg 100 µg 100 µg 100 µg 100 µg 100 µg 100 µg 100 µg 100 µg 100 µg 100 µg Contact Information Further information on Molecular Probes' products, including product bibliographies, is available from your local distributor or directly from Molecular Probes. Customers in Europe, Africa and the Middle East should contact our office in Leiden, the Netherlands. All others should contact our Technical Assistance Department in Eugene, Oregon. Please visit our Web site www.probes.com for the most up-to-date information Molecular Probes, Inc. 29851 Willow Creek Rd., Eugene, OR 97402 Phone: (541) 465-8300 Fax: (541) 344-6504 Molecular Probes Europe BV PoortGebouw, Rijnsburgerweg 10 2333 AA Leiden, The Netherlands Phone: +31-71-5233378 Fax: +31-71-5233419 Customer Service: 7:00 am to 5:00 pm (Pacific Time) Phone: (541) 465-8338 Fax: (541) 344-6504 [email protected] Customer Service: 9:00 to 16:30 (Central European Time) Phone: +31-71-5236850 Fax: +31-71-5233419 [email protected] Toll-Free Ordering for USA and Canada: Order Phone: (800) 438-2209 Order Fax: (800) 438-0228 Technical Assistance: 9:00 to 16:30 (Central European Time) Phone: +31-71-5233431 Fax: +31-71-5241883 [email protected] Technical Assistance: 8:00 am to 4:00 pm (Pacific Time) Phone: (541) 465-8353 Fax: (541) 465-4593 [email protected] Molecular Probes products are high-quality reagents and materials intended for research purposes only. These products must be used by, or directly under the supervision of, a technically qualified individual experienced in handling potentially hazardous chemicals. Please read the Material Safety Data Sheet provided for each product; other regulatory considerations may apply. Several of Molecular Probes products and product applications are covered by U.S. and foreign patents and patents pending. Our products are not available for resale or other commercial uses without a specific agreement from Molecular Probes, Inc. We welcome inquiries about licensing the use of our dyes, trademarks or technologies. Please submit inquiries by e-mail to [email protected]. All names containing the designation ® are registered with the U.S. Patent and Trademark Office. Copyright 2002, Molecular Probes, Inc. All rights reserved. This information is subject to change without notice. 6 Monoclonal Antibodies Specific for Oxidative Phosphorylation
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