Human/Mouse UCP1 Antibody

Detection of UCP1 in 3T3-L1 Mouse Cell Line by Flow Cytometry.

3T3-L1 mouse embryonic fibroblast adipose-like cell line was stained with Mouse Anti-Human/Mouse UCP1 Monoclonal Antibody (Catalog # MAB6158, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram) followed by anti-Mouse IgG PE-conjugated Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Detection of Mouse UCP1 by Simple Western^TM.

Simple Western lane view shows lysates of mouse adipose tissue and mouse brown adipose tissue, loaded at 0.5 mg/mL. A specific band was detected for UCP1 at approximately 37 kDa (as indicated) using 2.5 µg/mL of Mouse Anti-Human/Mouse UCP1 Monoclonal Antibody (Catalog # MAB6158). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Mouse UCP1 by Immunohistochemistry

IRX3/Irx3 expression is induced in browning adipose tissues.(A) Relative mRNA levels of Ucp1 and Irx3 in IWAT from mice under cold stress (4 °C) for one week (n = 6) and at 25 °C (n = 8). (B–C) Relative mRNA levels of Ucp1 and Irx3 in IWAT (B) and BAT (C) from mice subjected to PBS or CL-316,243 for 10 days. For A–C, gene expression was normalized to 36b4. (D–E) Protein levels of Ucp1 and Irx3 (left) and the quantification value of Irx3 relative to Hsp90 (right) in IWAT (D) and BAT (E) in the mice subjected to PBS or CL-316,243 treatment. (F) Representative images of immunohistochemical staining in IWAT from mice subjected to PBS or CL-316,243 treatment (scale bar, 50 μm for the upper and middle panels, and 100 μm for the bottom panel). (G) PET-CT, gross image, HE staining (scale bar, 20 μm), and electron microscopy (scale bar, 5 μm) of the browning WAT from pheochromocytoma patients. (H) Representative images of immunohistochemical staining in oWAT from pheochromocytoma patients (scale bar, 100 μm for the upper panel, and 50 μm for the bottom panel). Data were presented as mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001. The results are representative of at least three independent experiments. IWAT, inguinal white adipose tissue. BAT, brown adipose tissue. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28988979), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse UCP1 by Immunohistochemistry

IRX3/Irx3 expression is induced in browning adipose tissues.(A) Relative mRNA levels of Ucp1 and Irx3 in IWAT from mice under cold stress (4 °C) for one week (n = 6) and at 25 °C (n = 8). (B–C) Relative mRNA levels of Ucp1 and Irx3 in IWAT (B) and BAT (C) from mice subjected to PBS or CL-316,243 for 10 days. For A–C, gene expression was normalized to 36b4. (D–E) Protein levels of Ucp1 and Irx3 (left) and the quantification value of Irx3 relative to Hsp90 (right) in IWAT (D) and BAT (E) in the mice subjected to PBS or CL-316,243 treatment. (F) Representative images of immunohistochemical staining in IWAT from mice subjected to PBS or CL-316,243 treatment (scale bar, 50 μm for the upper and middle panels, and 100 μm for the bottom panel). (G) PET-CT, gross image, HE staining (scale bar, 20 μm), and electron microscopy (scale bar, 5 μm) of the browning WAT from pheochromocytoma patients. (H) Representative images of immunohistochemical staining in oWAT from pheochromocytoma patients (scale bar, 100 μm for the upper panel, and 50 μm for the bottom panel). Data were presented as mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001. The results are representative of at least three independent experiments. IWAT, inguinal white adipose tissue. BAT, brown adipose tissue. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28988979), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human UCP1 by Western Blot

BMP7 upregulated browning markers in both SC and DN derived differentiated adipocytes. SC and DN preadipocytes were differentiated and treated as in Figure 1. (A) Browning content as estimated by BATLAS Webtool (n = 9); (B) texture sum variance as quantified by laser-scanning cytometry (n = 5); quantification of UCP1 gene expression by (C) RNA-sequencing (n = 9) and (D) RT-qPCR normalized to GAPDH (n = 5). (E) UCP1 protein expression normalized to beta-actin (n = 5); (F) representative UCP1 immunostaining images visualized by confocal microscopy. Nucleus was stained using propidium iodide (PI), BF represents bright field image. Scale bars represent 10 μm. (G) Quantification of UCP1 immunostaining intensity assisted by laser-scanning cytometry (n = 4000 cells of 4 donors). Data expressed as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. Statistics: Paired t-test (A,B,G), GLM (C), and one-way ANOVA with Tukey’s post-test (D,E). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/34832860), licensed under a CC-BY license. Not internally tested by R&D Systems.