This antibody gave a positive signal in both Human Skeletal Muscle and Heart tissue lysates.
This antibody gave a positive result in IHC in the following FFPE tissue: Human normal Heart muscle.
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pH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 34 kDa).
UCP are mitochondrial transporter proteins that create proton leaks across the inner mitochondrial membrane, thus uncoupling oxidative phosphorylation. As a result, energy is dissipated in the form of heat. May play a role in the modulation of tissue respiratory control. Participates in thermogenesis and energy balance.
Only in skeletal muscle and heart. Is more expressed in glycolytic than in oxidative skeletal muscles.
Involvement in disease
Defects in UCP3 may be involved in obesity (OBESITY) [MIM:601665]. It is a condition characterized by an increase of body weight beyond the limitation of skeletal and physical requirements, as the result of excessive accumulation of body fat.
Belongs to the mitochondrial carrier family. Contains 3 Solcar repeats.
IHC image of UCP3 staining in a section of formalin-fixed paraffin-embedded normal Human heart muscle* performed on a Leica BONDTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab125830, 5ug/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.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Western blot - Anti-UCP3 antibody (ab125830)
All lanes : Anti-UCP3 antibody (ab125830) at 1 µg/ml
Lane 1 : Human skeletal muscle tissue lysate - total protein (ab29330) Lane 2 : Human heart tissue lysate - total protein (ab29431) Lane 3 : Liver (Human) Tissue Lysate - adult normal tissue (ab29889) - (Negative Control) Lane 4 : Kidney (Human) Tissue Lysate - adult normal tissue (ab30203)- (Negative Control)
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
The predicted molecular weight of UCP3 is 34 kDa (SwissProt), however we expect to observe a banding pattern between 45 and 50 kDa. Please note that both Human Liver and Kidney tissue lysates were used as negative controls for UCP3.
This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab125830 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
Giatromanolaki A et al. Thermogenic protein UCP1 and UCP3 expression in non-small cell lung cancer: relation with glycolysis and anaerobic metabolism. Cancer Biol Med14:396-404 (2017).
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