MitoBiogenesis™ Western Blot Cocktail (ab123545)
Overview
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Product name
MitoBiogenesis™ Western Blot Cocktail -
Species reactivity
Reacts with: Mouse, Rat, Cow, Human -
Product overview
The MitoBiogenesis™ western blotting cocktail (ab123545) is designed to study the regulation of mitochondrial biogenesis and cellular stress in response to environmental stimuli. It also can be used to monitor drug-induced effects on mitochondrial biogenesis. The two main components of this cocktail target two proteins, which are each subunits of a different oxidative phosphorylation enzyme complex, one subunit I of Complex IV (COX-I), which is mtDNA-encoded, and the 70kDa subunit of Complex II (SDH-A), which is nDNA-encoded. Complex IV includes several proteins which are encoded in the mitochondrion, while the proteins of Complex II are entirely encoded in the nucleus. An anti-Beta actin antibody is used as loading control.
Cocktail Antibodies:
Mouse CII-70 (SDHA) [2E3GC12FB2AE2] monoclonal:
Amount: 25 µg
Working concentration: 0.5 µg/ml
Mouse CIV-1 (MT-CO1) [1D6E1A8] monoclonal:
Amount: 50 µg
Working concentration: 1 µg/ml
Mouse Beta actin [mAbcam 8224] monoclonal:
Amount: 25 µg
Working concentration: 0.5 µg/ml
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Notes
Antibody Purity: Near homogeneity as judged by SDS-PAGE. The antibodies were produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation.
Related products
Review the mitochondrial assay guide, or the full metabolism assay guide 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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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 100 µg Antibody Cocktail 1 x 200µl -
Research areas
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Database links
- SwissProt: P23219 Human
- SwissProt: P60709 Human
- SwissProt: P31040 Human
Associated products
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Positive Controls
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab123545 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
Use at an assay dependent concentration.
The antibody cocktail should be diluted 250 X to a final working concentration of 2µg/mL for western blotting. COXI is a highly hydrophobic protein and appears as a broad band at ~35 kDa (not at its true molecular weight at 57 kDa). It is very sensitive to heating. Therefore, the samples, including the positive control, should not be heated over 50°C before loaded on the gel. |
Notes |
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WB
Use at an assay dependent concentration. The antibody cocktail should be diluted 250 X to a final working concentration of 2µg/mL for western blotting. COXI is a highly hydrophobic protein and appears as a broad band at ~35 kDa (not at its true molecular weight at 57 kDa). It is very sensitive to heating. Therefore, the samples, including the positive control, should not be heated over 50°C before loaded on the gel. |
Images
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All lanes : MitoBiogenesis™ Western Blot Cocktail (ab123545) at 1/250 dilution
Lane 1 : Human HepG2 cell lysate
Lane 2 : Human Hela cell lysate
Lane 3 : Human HDFn cell lysate
Lane 4 : Human SH-SY5Y cell lysate
Lane 5 : Rat H9C2 cell lysate
Lane 6 : Rat H4IIE cell lysate
Lane 7 : Mouse 3T3 cell lysate
Lane 8 : Mouse MEF cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : GAM-HRP at 1/3000 dilution -
All lanes : MitoBiogenesis™ Western Blot Cocktail (ab123545) at 1/250 dilution
Lane 1 : Human HepG2 cell lysate (vehicle treated)
Lane 2 : Human HepG2 cell lysate (Chloramphenicol treated)
Lane 3 : Human HepG2 cell lysate (DDC treated)
Lane 4 : Human 143B WT cell lysate
Lane 5 : Human 143B p0 cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : GAM-HRP at 1/3000 dilution
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (6)
ab123545 has been referenced in 6 publications.
- Nguyen TTT et al. Inhibition of HDAC1/2 Along with TRAP1 Causes Synthetic Lethality in Glioblastoma Model Systems. Cells 9:N/A (2020). PubMed: 32664214
- Nguyen TTT et al. HDAC inhibitors elicit metabolic reprogramming by targeting super-enhancers in glioblastoma models. J Clin Invest 130:3699-3716 (2020). PubMed: 32315286
- Rodríguez-García ME et al. A novel de novo MTOR gain-of-function variant in a patient with Smith-Kingsmore syndrome and Antiphospholipid syndrome. Eur J Hum Genet 27:1369-1378 (2019). PubMed: 31053780
- Gurley JM et al. Enhanced GLUT4-Dependent Glucose Transport Relieves Nutrient Stress in Obese Mice Through Changes in Lipid and Amino Acid Metabolism. Diabetes 65:3585-3597 (2016). WB ; Mouse . PubMed: 27679559
- Kiskinis E et al. Pathways disrupted in human ALS motor neurons identified through genetic correction of mutant SOD1. Cell Stem Cell 14:781-95 (2014). WB ; Human . PubMed: 24704492
- Zhou Y et al. Aristolochic acid causes albuminuria by promoting mitochondrial DNA damage and dysfunction in podocyte. PLoS One 8:e83408 (2013). WB ; Mouse . PubMed: 24349506