Monitoring of PPAR alpha protein expression in human tissue by the use of PPAR alpha-specific MAbs.
We report the production and characterization of two PPAR alpha subtype-specific monoclonal antibodies raised against the N-terminal domain of PPAR alpha. P alpha b 11.80A is a Western-reactive antibody, whereas P alpha b 32.51 is useful for immunohistochemistry. Both antibodies exhibited high affinity against the immunogen based on BIAcore analysis, recognized full-length PPAR alpha protein in PPAR alpha-transfected CV-1 cells, and displayed no cross-reactivity against the N-terminal domains of PPAR gamma or PPAR delta proteins as demonstrated by various immunoassays. The application of these antibodies to a panel of normal human tissues revealed that PPAR alpha protein expression is highest in skeletal muscle, liver, and kidney, consistent with previously reported mRNA expression data. These antibodies provide us with valuable tools to further explore the function of PPAR alpha.
Su
J L
JL
Department of Molecular Sciences, Glaxo Wellcome Research and Development, Research Triangle Park, NC 27709, USA.
Simmons
C J
CJ
Wisely
B
B
Ellis
B
B
Winegar
D A
DA
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D015374:Biosensing Techniques; D015153:Blotting, Western / Q000379:methods; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D009687:Nuclear Proteins / Q000032:analysis* / Q000276:immunology; D011487:Protein Conformation; D018160:Receptors, Cytoplasmic and Nuclear / Q000032:analysis* / Q000276:immunology; D014157:Transcription Factors / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D009687:Nuclear Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D014157:Transcription Factors