Quantitative expression patterns of peroxisome proliferator-activated receptor-β/δ (PPARβ/δ) protein in mice
Section snippets
Materials and methods
Anti-PPARβδ antibody characterization. Antiserum from six rabbits (designated 8095–8100) immunized with a PPARβ/δ-specific peptide (amino acids 1–29; sequence NP-035275) was obtained from Affinity Bioreagents. Antiserum was used to screen for PPARβ/δ immunoreactivity using Western blot analysis. COS-1 cells were transfected with a mouse PPARβ/δ expression vector, kindly provided by Drs. Walter Wahli and Pallavi Devchand, and cell lysate used for a positive control for these experiments. COS-1
Results
All six of antisera obtained from Affinity Bioreagents were immunoreactive with the COS-1 cell lysate with high PPARβ/δ expression (Fig. 1A). Of the six antisera examined, rabbit sera 8095, 8099, and 8100 were selected for further characterization after affinity purification. All three affinity purified antibodies immunoprecipitated in vitro-translated PPARβ/δ, with the percentage of immunoprecipitated protein ranging from 12–32% (Fig. 1B), which is quite efficient relative to other antibodies.
Discussion
Previous studies by others have described the expression patterns of PPARβ/δ mRNA in rat and mouse tissues with some variability noted. For example, one of the first studies that examined expression of PPARβ/δ mRNA by Northern blot analysis of rat samples noted the highest expression in adrenal gland, heart, and intestine, with modest expression in brain, kidney, and spleen, and low expression in liver and testis [10]. In mice, expression of PPARβ/δ mRNA was highest in mouse liver, with modest
Acknowledgments
Supported in part by the National Institutes of Health Grants CA97999 and CA124533 (J.M.P.).
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