Loading...
You have changed your country from
to
. Please be aware that this will change the currency in the purchasing process.
Related products
- Primary antibodies
- Lysates
Reassurance, Refunds & Replacements
If your product does not perform as described on this datasheet, we will refund or replace your product...
Read our guarantee »Overview
Product name
MitoProfile® Total OXPHOS Blue Native WB Antibody Cocktail
Assay type
Quantitative
Cross reactivity
Reacts with
Mouse, Rat, Cow, Human
Product overview
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 ab14713(MS111), CII-70 kDa subunit ab14715(MS204), CIII-Core protein 2 ab14745(MS304), CIV-subunit IV ab14744(MS407) and CV-alpha subunit 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 ab110240(MS103) and anti-20 kDa ab110242(MS105). A sample of purified 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 ab110247 (MS111), ab14715 (MS204), ab14745 (MS304), ab14744(MS407) and ab14748 (MS507).
Cocktail Antibodies:
Mouse monoclonal [20C11B11B11] to (C-I) NDUFA9 (ab14713):
Amount: 120 µg
Working Concentration: 2 µg/ml
Mouse monoclonal [2E3GC12FB2AE2] to (C-II-70) SDHA (ab14715):
Amount: 6 µg
Working Concentration: 0.1 µg/ml
Mouse monoclonal [13G12AF12BB11] to (C-III-Core 2) UQCRC2 (ab14745):
Amount: 60 µg
Working Concentration: 1 µg/ml
Mouse monoclonal [20E8C12] to (C-IV-subunit IV) COX IV(ab14744):
Amount: 60 µg
Working Concentration: 1 µg/ml
Mouse monoclonal [15H4C4 ] to (C-V-alpha) ATP5A (ab14748):
Amount: 60 µg
Working Concentration: 1 µg/ml
Notes
Store cocktail at 4°C and control at -80°C.
Tested applications
Properties
Storage instructions
Please see Notes section
| Components | 300 µg | 600 µg |
|---|---|---|
| Antibody Cocktail of 5 components | 1 x 300µg | 1 x 600µg |
| ab110338 - Bovine heart mitochondria control | 1 unit | 1 unit |
-
Western blot - MitoProfile® Total OXPHOS Blue Native WB Antibody Cocktail (ab110412)
(enlarge)
Alternative names
- MS603-300
Applications
Show applications key
Our Abpromise guarantee covers the use of ab110412 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: Use a concentration of 6 µg/ml. The antibody cocktail (1.5 mg/mL) should be diluted 250x to a final working concentration of 6.0 µg/mL for Western blotting
MitoProfile® Total OXPHOS Blue Native WB Antibody Cocktail images:
Western blot - MitoProfile® 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.
References for MitoProfile® Total OXPHOS Blue Native WB Antibody Cocktail (ab110412)
This product has been referenced in:
- Campanella M et al. Regulation of mitochondrial structure and function by the F1Fo-ATPase inhibitor protein, IF1. Cell Metab 8:13-25 (2008).Read more (PubMed: 18590689) »
- Kovárová N et al. Adaptation of respiratory chain biogenesis to cytochrome c oxidase deficiency caused by SURF1 gene mutations. Biochim Biophys Acta 1822:1114-1124 (2012).Read more (PubMed: 22465034) »
See all 3 publications for this product
Publishing research using ab110412? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"