WB: HLH, HHH, HeLa, HDFn, SY5Y, Jurkat, MLH, MHH, RLH, RHH, RBH, RKH, RSMH tissue homogenates
ICC/IF: Paraformaldehyde fixed HDFn (Human), NIH 3T3 (Mouse) and H9C2 (Rat) cells
Flow Cyt: HeLa cells
This antibody gave a positive result in IHC in the following FFPE tissue: Human normal heart muscle.
Product was previously marketed under the MitoSciences sub-brand.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use a concentration of 1 µg/ml. (cells fixed and permeabilized with methanol)
ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Use a concentration of 5 µg/ml. (paraformaldehyde fixed cells)
Use a concentration of 1 µg/ml. Predicted molecular weight: 68 kDa.
Use a concentration of 5 µg/ml.
Accepts electrons from ETF and reduces ubiquinone.
Involvement in disease
Defects in ETFDH are the cause of glutaric aciduria type 2C (GA2C) [MIM:231680]. GA2C is an autosomal recessively inherited disorder of fatty acid, amino acid, and choline metabolism. It is characterized by multiple acyl-CoA dehydrogenase deficiencies resulting in large excretion not only of glutaric acid, but also of lactic, ethylmalonic, butyric, isobutyric, 2-methyl-butyric, and isovaleric acids.
Belongs to the ETF-QO/fixC family. Contains 1 4Fe-4S ferredoxin-type domain.
Western blot - Anti-ETFDH antibody [3D1AC4AF3] (ab131376)
All lanes : Anti-ETFDH antibody [3D1AC4AF3] (ab131376) at 1 µg/ml
Lane 1 : Molecular weight marker Lane 2 : Human Liver Homogenate (HLH) at 10 µg Lane 3 : Human Heart Homogenate (HHH) at 10 µg Lane 4 : HepG2 Cell Lysate at 15 µg Lane 5 : Hela Cell Lysate at 15 µg Lane 6 : HDFn Cell Lysate at 15 µg Lane 7 : SY5Y Cell Lysate at 15 µg Lane 8 : Jurkat Cell Lysate at 15 µg Lane 9 : Mouse Liver Homogenate (MLH) at 10 µg Lane 10 : Mouse Heart Homogenate (MHH) at 10 µg Lane 11 : Rat Liver Homogenate (RLH) at 10 µg Lane 12 : Rat Heart Homogenate (RHH) at 10 µg Lane 13 : Rat Brain Homogenate (RBH) at 10 µg Lane 14 : Rat Kidney Homogenate (RLH) at 10 µg Lane 15 : Rat Skeletal Muscle Homogenate (RSMH) at 10 µg
Immunofluorescent analysis of ETFDH in HDFn cells.
The cells were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15 minutes). The cells were then incubated with ab131376 at 5 µg/ml for 2 h at room temperature, or over night at 4°C. The secondary antibody was (red) Alexa Fluor® 594 Goat anti-Mouse IgG (H+L) used at a 1/1000 dilution for 1 h. 1% BSA was used as the blocking agent for all blocking steps. The target protein locates to the mitochondria.
IHC image of ETFDH staining in Human normal heart muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab131376, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Flow cytometric analysis of ETFDH in HeLa cells fixed and permeabilized with methanol and stained with 1 µg/mL of ab131376 (blue), or an equal amount of an isotype control antibody (red). 1% BSA was used as the blocking reagent for all the blocking steps.
References for Anti-ETFDH antibody [3D1AC4AF3] (ab131376)
This product has been referenced in:
Roselló-Lletí E et al. Heart mitochondrial proteome study elucidates changes in cardiac energy metabolism and antioxidant PRDX3 in human dilated cardiomyopathy. PLoS One9:e112971 (2014).
Read more (PubMed: 25397948) »