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UCP-1 Antibody Staining Protocol for Immunohistochemistry
Description: The uncoupling protein UCP1 (formerly designated UCP) is an integral membrane protein unique to brown adipose tissue mitochondria. UCP1 forms a dimer that acts as a proton channel, which can uncouple oxidative phosphorylation by dissipating the electrochemical potential across the inner mitochondrial membrane. This process induces heat production in brown adipose tissue and is involved in regulation of body temperature and glucose metabolism. UCP2 is a structurally related protein that also uncouples mitochondrial respiration. It is more widely expressed in human and mouse tissues, including white adipose tissue and muscle, than is UCP1. UCP2 is thought to play a role in body weight regulation.
Primary Antibody
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Name: UCP-1 (C-17) Antibody |
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Clone: Goat polyclonal |
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Supplier: Santa Cruz Biotechnology |
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Catalog Number: sc-6528 |
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Dilution: 1:2000 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed. |
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Incubation Time/Temp: 60 min/room temperature |
Antigen Retrieval
| Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102) |
| Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100) |
| Heat/Cool Temperature: 95-100 ºC/room temperature |
| Heat/Cool Time: 20 minutes/20 minutes |
Detection Methods
| Standard Method: ABC Method or LSAB Method |
| Enhanced Method: Polymeric Methods |
Chromogen Substrate
| Reagent: DAB |
| Incubation Time/Temperature: 1-3 minutes/room temperature |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Cytoplasmic/membrane |
| Images: Search image |
Additional Information:
| Species Reactivity: Human, mouse, rat |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Pancreas (Islet cells) |
| Negative Control: Omit primary antibody, isotype control, absorption control |
| Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary |
References: