Tissue lysate was obtained by the total protein extraction kit (Solarbio, Beijing, China). After protein samples transferring to PVDF membranes, we cut off the extra PVDF membrane according to the molecular weight of the target protein. Then, the remaining PVDF membrane was incubated with a primary antibody. Antibodies were diluted 1:500 for rabbit anti-UCP1 (Cat: 72,298; Cell Signaling Technology, MA, USA), 1:500 for rabbit anti-PGC1α (Cat: A12348; ABclonal, Wuhan, China), 1:1000 for rabbit anti-β-Tubulin (Cat: AC008; Abclonal, Wuhan, China), and 1:1000 for HPR-labeled goat anti-rabbit IgG (Beyotime, Shanghai, China). Finally, a ChemiDoc Imaging Systems (Bio-Rad, CA, USA) was used to detect immunoreactive proteins.
Rabbit anti ucp1
Manufactured by Cell Signaling Technology
Sourced in United States, Germany
Rabbit anti-UCP1 is a primary antibody that specifically recognizes the Uncoupling Protein 1 (UCP1). UCP1 is a mitochondrial membrane protein that plays a crucial role in thermogenesis and energy expenditure.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti β tubulin, by ABclonal (2 mentions)
Mcp 1 ccl2, by R&D Systems (2 mentions)
Mab1501, by Merck Group (2 mentions)
Mouse anti mcp 1, by Thermo Fisher Scientific (2 mentions)
Mouse npy, by Merck Group (2 mentions)
Ma5 17040, by Thermo Fisher Scientific (2 mentions)
Rabbit anti gapdh, by Cell Signaling Technology (2 mentions)
Horse anti mouse, by Cell Signaling Technology (2 mentions)
Chemidoc imaging system, by Bio-Rad (1 mentions)
Hpr labeled goat anti rabbit igg, by Beyotime (1 mentions)
Total protein extraction kit, by Solarbio (1 mentions)
Triton x 100, by Solarbio (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Detergent compatible protein assay kit, by Bio-Rad (1 mentions)
Complete protease inhibitor cocktail, by Roche (1 mentions)
Mouse anti actin, by Merck Group (1 mentions)
Goat anti rabbit igg conjugated to horseradish peroxidase, by Cell Signaling Technology (1 mentions)
Ecl detection system, by Beyotime (1 mentions)
Protein extraction kit, by BestBio (1 mentions)
6 protocols using rabbit anti ucp1
1
Western Blot Analysis of UCP1 and PGC1α
2
Protein Extraction and Western Blotting
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Protein extraction and Western blotting were performed as described in [32 (link)] by applying the following primary antibodies: mouse anti-actin (MAB1501, 1:10,000, Merck Millipore); rabbit anti-Fhl2 (provided by the Schüle Laboratory, University of Freiburg, Freiburg im Breisgau, Germany); rabbit anti-GAPDH (#2118, 1:1000, Cell Signaling); mouse anti-MCP-1 (MA5-17040, 1:1000, Invitrogen); and rabbit anti-UCP1 (#14640, 1:1000, Cell Signaling). Mouse anti-rabbit (sc-2357, 1:10,000, Santa Cruz Biotechnology, Dallas, TX, USA) and horse anti-mouse (#7076, 1:3000, Cell Signaling) were used as secondary antibodies.
For the quantification of NPY and MCP-1 in tissue and cell protein lysates or supernatants, commercially available mouse NPY (Merck, Darmstadt, Germany) and CCL2/MCP-1 (R&D Systems, Minneapolis, MN, USA) ELISA kits were used according to the manufacturer’s instructions.
For the quantification of NPY and MCP-1 in tissue and cell protein lysates or supernatants, commercially available mouse NPY (Merck, Darmstadt, Germany) and CCL2/MCP-1 (R&D Systems, Minneapolis, MN, USA) ELISA kits were used according to the manufacturer’s instructions.
3
Protein Extraction and Western Blotting
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Protein extraction and Western blotting were performed as described in [32 (link)] by applying the following primary antibodies: mouse anti-actin (MAB1501, 1:10,000, Merck Millipore); rabbit anti-Fhl2 (provided by the Schüle Laboratory, University of Freiburg, Freiburg im Breisgau, Germany); rabbit anti-GAPDH (#2118, 1:1000, Cell Signaling); mouse anti-MCP-1 (MA5-17040, 1:1000, Invitrogen); and rabbit anti-UCP1 (#14640, 1:1000, Cell Signaling). Mouse anti-rabbit (sc-2357, 1:10,000, Santa Cruz Biotechnology, Dallas, TX, USA) and horse anti-mouse (#7076, 1:3000, Cell Signaling) were used as secondary antibodies.
For the quantification of NPY and MCP-1 in tissue and cell protein lysates or supernatants, commercially available mouse NPY (Merck, Darmstadt, Germany) and CCL2/MCP-1 (R&D Systems, Minneapolis, MN, USA) ELISA kits were used according to the manufacturer’s instructions.
For the quantification of NPY and MCP-1 in tissue and cell protein lysates or supernatants, commercially available mouse NPY (Merck, Darmstadt, Germany) and CCL2/MCP-1 (R&D Systems, Minneapolis, MN, USA) ELISA kits were used according to the manufacturer’s instructions.
4
Immunocytochemistry Analysis of Adipocytes
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We conducted immunocytochemistry analysis on brown and white adipocytes. The adipocytes were initially fixed with 4% paraformaldehyde for 15 min and subsequently incubated with 0.25% Triton X-100 (Solarbio, Beijing, China) at room temperature for 10 min. Subsequently, the washed cells were incubated overnight at 4 °C with the following primary antibodies: rabbit anti-UCP1 (dilution 1:100, Cell Signaling Technologies, Danvers, MA, USA) and rabbit anti-FGF11 (dilution 1:250, Bioss, Beijing, China). The cells were incubated with the secondary antibody (Cy3 Goat Anti-Rabbit IgG) diluted at 1:500 and maintained at 37 °C for 1 h. Finally, the cells were stained with DAPI and observed under a microscope.
5
Adipose Tissue Protein Analysis
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The adipose tissues were lysed in RIPA buffer containing 0.5% NP-40, 0.1% sodium deoxycholate, 150 mM NaCl, 50 mM Tris-Cl, pH 7.5, protease inhibitor cocktail (Complete, Roche), and 1 mM phenylmethylsulfonyl fluoride. Protein concentrations were quantified using a detergent-compatible protein assay kit (Bio-Rad). Lysates were run on Bis-Tris gels, transferred to a PVDF membrane (Millipore), and probed with primary antibodies. The following antibodies were used: rabbit anti-UCP1 (Cell Signaling, 72298), mouse anti-Actin (Millipore, MAB1501), and mouse anti-PGC1α (Sigma, ABE868). Goat-anti-rabbit IgG conjugated to horseradish peroxidase (Cell Signaling Technology) was used as the secondary antibody.
6
Protein Extraction and Western Blot Analysis
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The proteins used in the experiments were extracted using the protein extraction kit (Bestbio, Shanghai, China). The following primary antibodies were used: dilution 1:1000 for rabbit anti-UCP1 (Cell Signaling Technologies, MA, USA), and dilution 1:3000 for rabbit anti-β-Tubulin (Abclonal, Wuhan, China). The primary antibodies were incubated at 4 °C overnight. HRP-labeled goat anti-rabbit secondary antibody was diluted at 1:1000 for 1.5 h at 37 °C. Finally, the proteins were detected by the ECL detection system (Beyotime, Shanghai, China).
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