MS603 MitoProfile® Blue Native OXPHOS
Complexes Detection Kit
300 µg Kit: $575.00
|
|
600 µg Kit: $945.00
|
|
Please contact us for volume discounts.
Brief Description & Uses
Blue Native polyacrylamide gel electrophoresis, BN-PAGE (1), 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.
For a detailed protocol for BN-PAGE click here.
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 V can
be detected by mAbs against CI-NDUFA9 (MS111), CII-70 kDa subunit (MS204), CIII-Core protein 2 (MS304), CIV-subunit IV (MS407) and CV-α subunit (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 (MS103) and anti-20 kDa (MS105).
A sample of purified bovine heart mitochondria can be supplied with
these antibodies to act as a BNPAGE control sample upon request.
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. MitoSciences provides an
optimized pre-mixed cocktail of the mAbs to SIMULTANEOUSLY detect
Complexes I-V after 2nd dimension blotting (specifically the cocktail
contains MS111, MS204, MS304, MS407 and MS507). An example of this kind
of analysis in fibroblasts is shown in the figure below, also see
examples of the use of BN-PAGE in patient case studies (link below).
For a detailed description of 1st and 2nd dimension BN-PAGE click here. This cocktail is supplied with a sample of bovine heart mitochondria to act as a BNPAGE control sample.
Click here to review case studies for this product.
Specifications
Product:
|
MitoProfile® Blue Native OXPHOS Complexes
Detection Kit
|
Kit Components:
|
Premixed 306 µg cocktail containing the mAbs MS111, MS204, MS304, MS407 and MS507 (see Table below). BHM Control.
|
Catalog #:
|
MS603
|
Lot #:
|
|
Host Species:
|
Mouse
|
Concentration:
|
1.5 mg/ml in HEPES-Buffered Saline (HBS) with 0.02% azide as a preservative.
|
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.
|
Suggested Working Concentration:
|
The antibody cocktail (1.5 mg/ml) should be diluted 250x to a final working concentration of 6 µg/ml for Western blotting
|
Applications:
|
Western blotting detection OXPHOS complex assembly after BNPAGE.
|
Antigen Species Cross-Reactivity:
|
human, bovine and mouse
|
Storage:
|
4°C
|
Country of Origin:
|
USA
|
Kit Components
ANTIGEN
|
CAT #
|
CLONE ID
|
AMOUNT
(µg)
|
FINAL CONC.
(µg/ml)
|
IMMUNOGEN
|
C-I-39 (NDUFA9)
|
MS111
|
20C11B11B11
|
120
|
2
|
Bovine Heart Complex I
|
C-II-70 (Fp)
|
MS204
|
2E3GC12FB2AE2
|
6
|
0.1
|
Bovine Heart Complex II
|
C-III-Core 2
|
MS304
|
13G12AF12BB11
|
60
|
1
|
Bovine Heart Complex II, III
|
COXIV
|
MS407
|
20E8C12
|
60
|
1
|
Bovine Complex IV, holoenzyme
|
C-V-α
|
MS507
|
15H4C4
|
60
|
1
|
Bovine Complex V
|
Product References
There are no references currently posted for this product.
Have you published research using this product?
Let us know and earn a $100.00 discount on your next purchase.
References for Oxidative Phosphorylation
There are no references currently posted for this product.
Images
|
 |
Downloadable Information |
|
|
|
|
|