Mouse Monoclonal ACADM/MCAD antibody. Suitable for IHC-P, Flow Cyt, WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 12 publications.
pH: 7.5
Preservative: 0.02% Sodium azide
Constituents: 99.98% HEPES buffered saline
IHC-P | Flow Cyt | WB | ICC/IF | |
---|---|---|---|---|
Human | Tested | Tested | Tested | Tested |
Mouse | Expected | Expected | Tested | Expected |
Rat | Expected | Expected | Tested | Expected |
Species | Dilution info | Notes |
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Species Human | Dilution info 1/1000 | Notes Perform heat-mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1 µg/mL | Notes ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 0.125 µg/mL | Notes - |
Species Mouse | Dilution info 0.125 µg/mL | Notes - |
Species Rat | Dilution info 0.125 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
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Medium-chain specific acyl-CoA dehydrogenase is one of the acyl-CoA dehydrogenases that catalyze the first step of mitochondrial fatty acid beta-oxidation, an aerobic process breaking down fatty acids into acetyl-CoA and allowing the production of energy from fats (PubMed:1970566, PubMed:21237683, PubMed:2251268, PubMed:8823175). The first step of fatty acid beta-oxidation consists in the removal of one hydrogen from C-2 and C-3 of the straight-chain fatty acyl-CoA thioester, resulting in the formation of trans-2-enoyl-CoA (PubMed:2251268). Electron transfer flavoprotein (ETF) is the electron acceptor that transfers electrons to the main mitochondrial respiratory chain via ETF-ubiquinone oxidoreductase (ETF dehydrogenase) (PubMed:15159392, PubMed:25416781). Among the different mitochondrial acyl-CoA dehydrogenases, medium-chain specific acyl-CoA dehydrogenase acts specifically on acyl-CoAs with saturated 6 to 12 carbons long primary chains (PubMed:1970566, PubMed:21237683, PubMed:2251268, PubMed:8823175).
MCAD, Medium chain acyl-CoA dehydrogenase, MCADH, ACADM
Mouse Monoclonal ACADM/MCAD antibody. Suitable for IHC-P, Flow Cyt, WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 12 publications.
pH: 7.5
Preservative: 0.02% Sodium azide
Constituents: 99.98% HEPES buffered saline
Produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation.
MCAD (medium-chain acyl-CoA dehydrogenase) is an oxidoreductase enzyme of the mitochondrial fatty acid beta-oxidation pathway that is specific for acyl chain lengths of 4 to 16. It also utilizes the electron transfer flavoprotein (ETF) as electron acceptor that transfers the electrons to the main mitochondrial respiratory chain via ETF-ubiquinone oxidoreductase (ETF dehydrogenase).
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ACADM also known as MCAD (medium-chain acyl-CoA dehydrogenase) is an enzyme that plays a significant role in fatty acid metabolism. It weighs approximately 44 kDa and expresses mainly in the liver heart and skeletal muscles. Functions of MCAD involve catalyzing the initial step in the mitochondrial fatty acid β-oxidation spiral specifically for the medium-chain fatty acids. This enzyme helps in the conversion of fatty acyl-CoA to trans-enoyl-CoA through dehydrogenation.
The enzyme facilitates energy production by breaking down medium-chain fatty acids within mitochondria. MCAD operates as a homotetramer complex where each subunit significantly contributes to its overall function. Such enzymatic activity is important for providing energy particularly when glycogen stores are low. Its efficient operation during fasting states indicates its importance in metabolic homeostasis.
Medium-chain acyl-CoA dehydrogenase (MCAD) is integral to the mitochondrial fatty acid beta-oxidation pathway. This pathway is a primary route for fatty acid catabolism ultimately leading to energy production in the form of ATP. MCAD's activity also relates to other fatty acid oxidation enzymes like VLCAD (very-long-chain acyl-CoA dehydrogenase) and SCAD (short-chain acyl-CoA dehydrogenase) which operate on different chain-length fatty acids cooperating to maintain energy balance.
MCAD deficiency is a common metabolic disorder that impairs the normal breakdown of fatty acids. This condition results in the accumulation of fatty acid intermediates leading to hypoketotic hypoglycemia during fasting periods. Individuals with MCAD deficiency may experience lethargy vomiting and seizures. The disorder connects with other enzymes such as LCHAD (long-chain 3-hydroxyacyl-CoA dehydrogenase) involved in similar pathways and deficiencies can result in related metabolic dysfunctions.
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All lanes: Western blot - Anti-ACADM/MCAD antibody [3B7BH7] (ab110296) at 0.125 µg/mL
Lane 1: HepG2 cell lysates at 15 µg
Lane 2: HeLa cell lysates at 15 µg
Lane 3: H9C2 (rat cells) lysates at 15 µg
Lane 4: MEF (mouse cells) lysates at 15 µg
Predicted band size: 47 kDa
HL-60 cells were stained with 1 µg/mL ab110296 (blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.
Immunocytochemistry image of ab110296 stained Human HeLa cells. The cells were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15 minutes). The cells were incubated with ab110296 (1 µg/ml) for 2 hours at room temperature or over night at 4°C. The secondary antibody was (green) Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1 hour. 10% Goat serum was used as the blocking agent for all blocking steps. DAPI was used to stain the cell nuclei (blue). Target protein locates mainly in mitochondria.
ACADM/MCAD immunohistochemistry in Human cerebellum visualized with ab110296 at 1/1000. MCAD immunoactivity is most intense in neuronal cell bodies, most notably in the large Purkinje cells. Note the distinctive subcellular localization of MCAD immunoreactivity in the Purkinje cell bodies.
HL-60 cells were incubated at 37°C for 24h with vehicle control (0 μM) and different concentrations of fenofibrate (Fenofibrate, PPAR-alpha agonist ab120832). Increased expression of ACADM/MCAD in HL-60 cells correlates with an increase in fenofibrate concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab110296 at 1 μg/ml and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 at 1 μg/ml overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP (Goat Anti-Mouse IgG H&L (HRP) preadsorbed ab97040) at 1/10000 dilution and visualised using ECL development solution.
All lanes: Western blot - Anti-ACADM/MCAD antibody [3B7BH7] (ab110296)
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 47 kDa
Image collected and cropped by CiteAb under a CC-BY license from the publication
ACADM/MCAD western blot using anti-ACADM/MCAD antibody [3B7BH7] ab110296. Publication image and figure legend from Gao, K., Zhang, J., et al., 2020, Chin Med, PubMed 32158496.
ab110296 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab110296 please see the product overview.
Effects of QSG on FAT/CD36-CPT1-FAO pathway in HF rats after AMI. The protein expressional levels of CD36, CPT1A, ACADL, ACADM, ACAA2 and SCP2 in the three groups were significantly decreased in the model group compared with the sham group. After treatment with QSG, cardiac CD36, CPT1A, ACADL, ACADM, ACAA2 and SCP2 levels were all significantly increased. n = 4 per group. Values are mean ± SE. Asterisks indicates significant differences. *P < 0.05, **P < 0.01
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