Western blot analysis of PPAR-alpha was performed by loading 20 µg of Mouse Liver (lane1), Jurkat (lane2), HeLa (lane3), NIH/3T3 (lane4) and PC-12 (lane5) lysate using Novex® NuPAGE® 4-12 % Bis-Tris gel (NP0321BOX), XCell SureLock™ Electrophoresis System (EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature. PPAR-alpha was detected at ~58 kDa using PPAR-alpha Rabbit Polyclonal Antibody (424600) at 1-3 µg/ml in 5% skim milk at 4°C overnight on a rocking platform. Goat Anti-Rabbit IgG - HRP Secondary Antibody (G21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (WP20005).
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||specific to Human PPAR alpha|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:100-1:500|
|Western Blot (WB)||1-3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Peroxisome proliferator-activated receptor alpha (PPARA) is a member of the nuclear hormone receptor subfamily. Upon activation, PPARA forms heterodimers with retinoid X receptors (RXRs), and functions to regulatetranscrption of specific gene sequences. In particular, PPARA controls genes involved in the peroxisomal and mitochondrial beta oxidation pathways, fatty acid uptake, and triglyceride catabolism in the liver and skeletal muscle. PPARA also has anti-inflammatory and anti-proliferative effects and prevents the proatherogenic effects of cholesterol accumulation in macrophages by stimulating cholesterol efflux. PPARA is the main target of fibrate drugs; hypolipidemic agents used in the treatment of dyslipidemia and the prevention of cardiovascular disease. They could also be used for the treatment of type II diabetes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.