Mouse Monoclonal ETFA antibody. Suitable for Flow Cyt, WB, ICC/IF, IHC-P and reacts with Human, Mouse, Rat, Cow samples. Cited in 8 publications.
IgG2b
Mouse
pH: 7.5
Preservative: 0.02% Sodium azide
Constituents: HEPES buffered saline
Liquid
Monoclonal
Flow Cyt | WB | ICC/IF | IHC-P | |
---|---|---|---|---|
Human | Tested | Tested | Tested | Tested |
Mouse | Expected | Tested | Expected | Expected |
Rat | Expected | Tested | Expected | Expected |
Cow | Expected | Tested | Expected | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1 µg/mL | Notes ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Cow | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1 µg/mL | Notes - |
Species Mouse | Dilution info 1 µg/mL | Notes - |
Species Rat | Dilution info 1 µg/mL | Notes - |
Species Cow | Dilution info 1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 4 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Cow | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 5 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Cow | Dilution info Use at an assay dependent concentration. | Notes - |
Select an associated product type
Heterodimeric electron transfer flavoprotein that accepts electrons from several mitochondrial dehydrogenases, including acyl-CoA dehydrogenases, glutaryl-CoA and sarcosine dehydrogenase (PubMed:10356313, PubMed:15159392, PubMed:15975918, PubMed:27499296, PubMed:9334218). It transfers the electrons to the main mitochondrial respiratory chain via ETF-ubiquinone oxidoreductase (ETF dehydrogenase) (PubMed:9334218). Required for normal mitochondrial fatty acid oxidation and normal amino acid metabolism (PubMed:12815589, PubMed:1430199, PubMed:1882842).
Alpha-ETF, ETFA
Mouse Monoclonal ETFA antibody. Suitable for Flow Cyt, WB, ICC/IF, IHC-P and reacts with Human, Mouse, Rat, Cow samples. Cited in 8 publications.
IgG2b
Mouse
pH: 7.5
Preservative: 0.02% Sodium azide
Constituents: HEPES buffered saline
Liquid
Monoclonal
2B11AE8
Proprietary technique
kappa
The antibody was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation. Purity >95% by SDS-PAGE.
Blue Ice
+4°C
+4°C
Do Not Freeze
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Product was previously marketed under the MitoSciences sub-brand.
This supplementary information is collated from multiple sources and compiled automatically.
ETFA or Electron-Transfer-Flavoprotein alpha subunit is an essential part of the mitochondrial respiratory chain. This protein which has a molecular mass of approximately 34 kDa functions in the transfer of electrons from acyl-CoA dehydrogenases to the main respiratory chain for energy production. ETFA is commonly expressed in tissues with high-energy demands such as the liver heart and skeletal muscle. The protein forms a heterodimeric complex with its counterpart ETFB providing a critical function in electron transfer during fatty acid oxidation.
ETFA operates as an important part of the electron transfer process within the mitochondria. It acts as one-half of the heterodimeric electron-transfer flavoprotein complex teaming with ETFB. This complex facilitates electron transfer from a range of acyl-CoA dehydrogenases to ETF dehydrogenase which then continues the process of electron transfer to coenzyme Q in the respiratory chain. This action is key to the breakdown of fats enabling energy extraction and processing.
ETFA has important roles in fatty acid beta-oxidation and amino acid catabolism. It engages in these pathways by transferring electrons as mentioned interfacing with other proteins like ETF dehydrogenase. This positioning within the mitochondrial matrix enables ETFA to assist in converting fat and protein substrates into energy which the cell can use. Its molecular interactions highlight its integral position in maintaining energy homeostasis.
Problems with ETFA lead to glutaric acidemia type 2 a metabolic disorder that impairs the body's ability to oxidize fatty acids and some amino acids. Deficiencies in ETFA function disrupt the electron transport to the respiratory chain causing an accumulation of intermediary metabolites. These disruptions can relate to or involve other mitochondrial components and proteins like ETFB or ETFDH. Correct diagnosis and understanding of ETFA’s role in such conditions are instrumental for targeted therapeutic approaches.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Lanes 1-2: Merged signal (red and green). Green - ab110316 observed at 35 kDa. Red - loading control Anti-beta Tubulin antibody [EP1331Y] - Microtubule Marker ab52901 observed at kDa.
ab110316 Anti-ETFA antibody [2B11AE8] was shown to specifically react with ETFA in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line Human ETFA knockout HEK-293T cell line ab266513 (knockout cell lysate Human ETFA knockout HEK-293T cell lysate ab257943) was used. Wild-type and ETFA knockout samples were subjected to SDS-PAGE. ab110316 and Anti-beta Tubulin [EP1331Y] - Microtubule Marker (Anti-beta Tubulin antibody [EP1331Y] - Microtubule Marker ab52901) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-ETFA antibody [2B11AE8] (ab110316) at 1/1000 dilution
Lane 1: Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 40 µg
Lane 2: ETFA knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 40 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/10000 dilution
Predicted band size: 35 kDa, 81 kDa
Observed band size: 35 kDa
HL-60 cells were stained with 1 µg/mL ab110316 (blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.
All lanes: Western blot - Anti-ETFA antibody [2B11AE8] (ab110316) at 1 µg/mL
Lane 1: Human heart tissue at 10 µg
Lane 2: HepG2 whole cells at 10 µg
Lane 3: Human liver mitochondria at 10 µg
Lane 4: Bovine heart mitochondria at 10 µg
Lane 5: Rat liver mitochondria at 10 µg
Lane 6: Mouse liver mitochondria at 10 µg
Predicted band size: 35 kDa
Immunocytochemistry image of stained HeLa cells. The cells were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15 minutes). The cells were incubated with the antibody (ab110316, 4 µg/mL) for 2 hours at room temperature or over night at 4°C. The secondary antibody was (red) Alexa Fluor® 594 goat anti-mouse IgG (H+L) at a 1/1000 dilution for 1 hour. 10% Goat serum was used as the blocking agent for all blocking steps. The target protein locates to the mitochondrial matrix
IHC image of ETFA staining in Human heart 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 ab110316, 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.
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