Total OXPHOS Blue Native WB Antibody Cocktail (ab110412) is part of the reagents, controls & accessories range. Abcam offers high-quality biological reagents and tools including antibodies, proteins, assays, cell lines and lysates.
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- Science advances 9:eadh05582023Cross-species analysis identifies mitochondrial dysregulation as a functional consequence of the schizophrenia-associated 3q29 deletion.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRyan H Purcell,Esra Sefik,Erica Werner,Alexia T King,Trenell J Mosley,Megan E Merritt-Garza,Pankaj Chopra,Zachary T McEachin,Sridhar Karne,Nisha Raj,Brandon J Vaglio,Dylan Sullivan,Bonnie L Firestein,Kedamawit Tilahun,Maxine I Robinette,Stephen T Warren,Zhexing Wen,Victor Faundez,Steven A Sloan,Gary J Bassell,Jennifer G MullePubMed 37585521
- International journal of molecular sciences 23:2022Desmin Knock-Out Cardiomyopathy: A Heart on the Verge of Metabolic Crisis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesBarbara Elsnicova,Daniela Hornikova,Veronika Tibenska,David Kolar,Tereza Tlapakova,Benjamin Schmid,Markus Mallek,Britta Eggers,Ursula Schlötzer-Schrehardt,Viktoriya Peeva,Carolin Berwanger,Bettina Eberhard,Hacer Durmuş,Dorothea Schultheis,Christian Holtzhausen,Karin Schork,Katrin Marcus,Jens Jordan,Thomas Lücke,Peter F M van der Ven,Rolf Schröder,Christoph S Clemen,Jitka M ZurmanovaPubMed 36233322
- American journal of physiology. Lung cellular and molecular physiology 323:L355-L3712022Contribution of fatty acid oxidation to the pathogenesis of pulmonary hypertension.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMichael H Lee,Linda Sanders,Rahul Kumar,Daniel Hernandez-Saavedra,Xin Yun,Joshay A Ford,Mario J Perez,Claudia Mickael,Aneta Gandjeva,Daniel E Koyanagi,Julie W Harral,David C Irwin,Biruk Kassa,Robert H Eckel,Larissa A Shimoda,Brian B Graham,Rubin M TuderPubMed 35763400
- Molecular metabolism 61:1014992022Loss of UCP1 function augments recruitment of futile lipid cycling for thermogenesis in murine brown fat.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJosef Oeckl,Petra Janovska,Katerina Adamcova,Kristina Bardova,Sarah Brunner,Sebastian Dieckmann,Josef Ecker,Tobias Fromme,Jiri Funda,Thomas Gantert,Piero Giansanti,Maria Soledad Hidrobo,Ondrej Kuda,Bernhard Kuster,Yongguo Li,Radek Pohl,Sabine Schmitt,Sabine Schweizer,Hans Zischka,Petr Zouhar,Jan Kopecky,Martin KlingensporPubMed 35470094
- Human molecular genetics 31:3597-36122022Deleterious variants in CRLS1 lead to cardiolipin deficiency and cause an autosomal recessive multi-system mitochondrial disease.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRichard G Lee,Shanti Balasubramaniam,Maike Stentenbach,Tom Kralj,Tim McCubbin,Benjamin Padman,Janine Smith,Lisa G Riley,Archana Priyadarshi,Liuyu Peng,Madison R Nuske,Richard Webster,Ken Peacock,Philip Roberts,Zornitza Stark,Gabrielle Lemire,Yoko A Ito,Kym M Boycott,Michael T Geraghty,Jan Bert van Klinken,Sacha Ferdinandusse,Ying Zhou,Rebecca Walsh,Esteban Marcellin,David R Thorburn,Tony Rosciolli,Janice Fletcher,Oliver Rackham,Frédéric M Vaz,Gavin E Reid,Aleksandra FilipovskaPubMed 35147173
- Journal of neurochemistry 160:540-5552022Drosophila ZIP13 over-expression or transferrin1 RNAi influences the muscle degeneration of Pink1 RNAi by elevating iron levels in mitochondria.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJinsong Xue,Guangying Li,Xiaowen Ji,Zhi-Hua Liu,Hui-Li Wang,Guiran XiaoPubMed 35038358
- Molecular biology of the cell 33:ar292022Ovarian carcinoma immunoreactive antigen-like protein 2 (OCIAD2) is a novel complex III-specific assembly factor in mitochondria.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKatarzyna Justyna Chojnacka,Praveenraj Elancheliyan,Ben Hur Marins Mussulini,Karthik Mohanraj,Sylvie Callegari,Aleksandra Gosk,Tomasz Banach,Tomasz Góral,Karolina Szczepanowska,Peter Rehling,Remigiusz Adam Serwa,Agnieszka ChacinskaPubMed 35080992
- Nature communications 13:1342022Mutations in Hcfc1 and Ronin result in an inborn error of cobalamin metabolism and ribosomopathy.Applications:Unspecified applicationReactive species:Unspecified reactive speciesTiffany Chern,Annita Achilleos,Xuefei Tong,Matthew C Hill,Alexander B Saltzman,Lucas C Reineke,Arindam Chaudhury,Swapan K Dasgupta,Yushi Redhead,David Watkins,Joel R Neilson,Perumal Thiagarajan,Jeremy B A Green,Anna Malovannaya,James F Martin,David S Rosenblatt,Ross A PochéPubMed 35013307
- Particle and fibre toxicology 18:442021Transcriptomics of single dose and repeated carbon black and ozone inhalation co-exposure highlight progressive pulmonary mitochondrial dysfunction.Applications:Unspecified applicationReactive species:Unspecified reactive speciesQuincy A Hathaway,Nairrita Majumder,William T Goldsmith,Amina Kunovac,Mark V Pinti,Jack R Harkema,Vince Castranova,John M Hollander,Salik HussainPubMed 34911549
- BMC biology 19:2652021Enhanced hepatic respiratory capacity and altered lipid metabolism support metabolic homeostasis during short-term hypoxic stress.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKatie A O'Brien,Ben D McNally,Alice P Sowton,Antonio Murgia,James Armitage,Luke W Thomas,Fynn N Krause,Lucas A Maddalena,Ian Francis,Stefan Kavanagh,Dominic P Williams,Margaret Ashcroft,Julian L Griffin,Jonathan J Lyon,Andrew J MurrayPubMed 34911556
Key facts
- Applications
- WB
- Reactive species
- Mouse, Rat, Cow, Human
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application WB | Reactivity Reacts | Dilution info - | Notes The antibody cocktail (1.5 mg/mL) should be diluted 250x to a final working concentration of 6.0 µg/mL for Western blotting |
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Total OXPHOS Blue Native WB Antibody Cocktail (ab110412) is part of the reagents, controls & accessories range. Abcam offers high-quality biological reagents and tools including antibodies, proteins, assays, cell lines and lysates.
Key facts
- Applications
- WB
- Reactive species
- Mouse, Rat, Cow, Human
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -80°C
- Appropriate long-term storage conditions
- -80°C
Notes
Blue Native polyacrylamide gel electrophoresis (BN-PAGE) is a simple and effective way to subfractionate mitochondrial proteins as intact complexes on a single gel (in one dimension). It can be used to detect altered assembly of these complexes arising from mutations in subunits, mutations in assembly factors, or mtDNA depletion. This type of analysis has been performed with biopsy samples, platelets and fibroblast cells from patients with suspected mitochondrial diseases.
In this method, multisubunit enzymes bind a charged dye Coomassie brilliant blue which allows their electrophoretic separation in the first dimension by the size of the complex. Complexes I-V with masses ranging from 950K to 200K are well resolved in the first dimension. The separated proteins can then be transferred to nitrocellulose membrane/PVDF by electrophoresis and Complexes I, II, III, IV and ATP synthase can be detected by mAbs against CI-NDUFA9 Anti-NDUFA9 antibody [20C11B11B11] ab14713(MS111), CII-70 kDa subunit Anti-SDHA antibody [2E3GC12FB2AE2] ab14715(MS204), CIII-Core protein 2 Anti-UQCRC2 antibody [13G12AF12BB11] ab14745(MS304), CIV-subunit IV Anti-COX IV antibody [20E8C12] - Mitochondrial Marker ab14744(MS407) and CV-alpha subunit Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker ab14748(MS507) respectively. Such one dimensional gels, are best analyzed by using single mAbs against each complex. Other Complex I mAbs are available for BNPAGE, specifically anti-GRIM-19 Anti-GRIM19 antibody [6E1BH7] ab110240(MS103) and anti-20 kDa Anti-NDUFB8 antibody [20E9DH10C12] ab110242(MS105). A sample of purified bovine heart mitochondria Bovine Heart Mitochondria ab110338(MS802) is available as a BNPAGE control sample.
Sometimes a greater separation of enzymes is necessary - it is possible to separate the proteins within each individual complex. To do this, blotting is NOT performed after the first (NATIVE) dimension, instead gels are turned 90 degrees and run in a perpendicular second dimension which is denaturing (NON-NATIVE). In this way the protein subunits within each complex are separated. Abcam provides an optimized pre-mixed cocktail of the mAbs to SIMULTANEOUSLY detect Complexes I-V after 2nd dimension blotting (specifically the cocktail contains Anti-NDUFA9 antibody [20C11B11B11] ab14713 (MS111), Anti-SDHA antibody [2E3GC12FB2AE2] ab14715 (MS204), Anti-UQCRC2 antibody [13G12AF12BB11] ab14745 (MS304), Anti-COX IV antibody [20E8C12] - Mitochondrial Marker ab14744(MS407) and Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker ab14748 (MS507).
Cocktail Antibodies:
Mouse monoclonal [20C11B11B11] to (C-I) NDUFA9 (Anti-NDUFA9 antibody [20C11B11B11] ab14713):
Amount: 120 μg
Working Concentration: 2 μg/ml
Mouse monoclonal [2E3GC12FB2AE2] to (C-II-70) SDHA (Anti-SDHA antibody [2E3GC12FB2AE2] ab14715):
Amount: 6 μg
Working Concentration: 0.1 μg/ml
Mouse monoclonal [13G12AF12BB11] to (C-III-Core 2) UQCRC2 (Anti-UQCRC2 antibody [13G12AF12BB11] ab14745):
Amount: 60 μg
Working Concentration: 1 μg/ml
Mouse monoclonal [20E8C12] to (C-IV-subunit IV) COX IV(Anti-COX IV antibody [20E8C12] - Mitochondrial Marker ab14744):
Amount: 60 μg
Working Concentration: 1 μg/ml
Mouse monoclonal [15H4C4 ] to (C-V-alpha) ATP5A (Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker ab14748):
Amount: 60 μg
Working Concentration: 1 μg/ml
**Related products**Review the , or the full to learn about more assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also how to assay metabolic function in live cells using your plate reader.
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Western blot - Total OXPHOS Blue Native WB Antibody Cocktail (ab110412)
Two dimension Blue Native PAGE analysis of fibroblasts that are (A) normal and (B) complex I deficient. It is clear that the complex I deficient cell line shown in B shows no detectable level of complex I. However, all other OXPHOS complexes appear normal including a minor amount of heterocomplex formed between complexes III and IV which is a well documented species. Only a minor mitochondrial enrichment was performed upon these samples cell lines. Each sample represents only 8% of confluent cells taken from a 10 cm diameter tissue culture dish.
MWs:
(C-I) NDUFA9 - 36kDa
(C-II-70) SDHA - 70kDA
(C-III-Core 2) UQRC2) - 45kDa
(C-IV subunit IV) COX IV - 15kDa
(C-V alpha) ATP5A - 55kDa
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