Thermogenin (called uncoupling protein by its discoverers and now known as uncoupling protein 1, or UCP1) is a mitochondrial carrier protein found in brown adipose tissue (BAT). It is used to generate heat by non-shivering thermogenesis, and makes a quantitatively important contribution to countering heat loss in babies which would otherwise occur due to their high surface area-volume ratio. Recent findings indicate that the UCP1 protein plays a crucial role in thermogenesis by catalyzing the dissipative production of heat through protons derived from NADH and FADH2. These electron carriers are produced in the TCA cycle from the oxidation of acetyl-CoA, which comes from the breakdown of free fatty acids. Intriguingly, the acetyl-CoA products undergo a recycling process that facilitates their re-utilization, thereby sustaining the cycle known as the HEAT cycle.
Background References
1. Rahbani JF et al. Parallel control of cold-triggered adipocyte thermogenesis by UCP1 and CKB. Cell Metab. 2024 Mar
2. Johnson JM et al. Mitochondrial phosphatidylethanolamine modulates UCP1 to promote brown adipose thermogenesis. Sci Adv. 2023 Feb
Western blot analysis of UCP1 on different lysates with Rabbit anti-UCP1 antibody (HA723776) at 1/2,000 dilution.
Lane 1: Mouse brown adipose tissue lysate Lane 2: Rat brown adipose tissue lysate Lane 3: Rat white adipose tissue lysate (negative)
Lysates/proteins at 20 µg/Lane.
Predicted band size: 33 kDa Observed band size: 33 kDa
Exposure time: 3 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723776) at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Application: IHC-Fr
Species: Mouse
Site: brown adipose
Sample: Frozen section
Antibody concentration: 1/500
Antigen retrieval: Not required
Immunohistochemical analysis of paraffin-embedded mouse brown adipose tissue with Rabbit anti-UCP1 antibody (HA723776) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723776) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
☑ Relative expression (RE)
Immunohistochemical analysis of paraffin-embedded mouse white adipose tissue (negative) with Rabbit anti-UCP1 antibody (HA723776) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723776) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded rat brown adipose tissue with Rabbit anti-UCP1 antibody (HA723776) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723776) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
☑ Relative expression (RE)
Immunohistochemical analysis of paraffin-embedded rat white adipose tissue (negative) with Rabbit anti-UCP1 antibody (HA723776) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723776) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
UCP1 was immunoprecipitated from 0.2 mg mouse brown adipose tissue lysate with HA723776 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723776 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: mouse brown adipose tissue lysate (input) Lane 2: HA723776 IP in mouse brown adipose tissue lysate Lane 3: Rabbit IgG instead of HA723776 in mouse brown adipose tissue lysate