Immunofluorescence analysis of PPAR alpha was done on 70% confluent log phase HCT116 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PPAR alpha (3B6/PPAR) Mouse Monoclonal Antibody (MA1822) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic and nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rabbit, Rat, Bovine, Mouse, Human|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Purified recombinant PPAR alpha protein.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||Assay Dependent|
|Flow Cytometry (Flow)||1µg per 10^6 cells|
|Gel Shift (GS)||Assay dependent|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||2-5 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.
MA1-822 detects peroxisome proliferator activated receptors (PPAR) alpha from human, mouse, and rat tissues.
MA1-822 has been successfully used in Western blot, Gel Shift and immunoprecipitation procedures. By Western blot, this antibody detects a ~52 kDa protein which corresponds to PPAR alpha from mouse adipose tissue extract. This antibody detects some non-specific bands on NIH-3T3 cell lysates.
The MA1-822 immunogen is purified recombinant PPAR alpha protein.
Peroxisome proliferators are non-genotoxic carcinogens which are purported to exert their effect on cells through their interaction with members of the nuclear hormone receptor family termed peroxisome proliferator activated receptors (PPAR and quote;s). Nuclear hormone receptors are ligand-dependent intracellular proteins that stimulate transcription of specific genes by binding to specific DNA sequences following activation by the appropriate ligand. Studies indicate that PPAR and quote;s are activated by peroxisome proliferators such as clofibric acid, nafenopin, and WY-14,643, as well as by some fatty acids. It has also been shown that PPAR and quote;s can induce transcription of acyl coenzyme A oxidase and cytochrome P450 (CYP450) A6 through interaction with specific response elements. PPAR, like several other nuclear hormone receptors, heterodimerizes with retinoid X receptor (RXR) alpha.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Selective reversible inhibition of liver carnitine palmitoyl-transferase 1 by teglicar reduces gluconeogenesis and improves glucose homeostasis.
MA1-822 was used in western blot to investigate the effect of liver carnitine palmitoyl-transferase 1 inhibition on gluconeogenesis and glucose homeostasis
|Conti R,Mannucci E,Pessotto P,Tassoni E,Carminati P,Giannessi F,Arduini A||Diabetes (60:644)||2011|
Noradrenaline represses PPAR (peroxisome-proliferator-activated receptor) gamma2 gene expression in brown adipocytes: intracellular signalling and effects on PPARgamma2 and PPARgamma1 protein levels.
MA1-822 was used in western blot to investigate the effect of noradrenaline on PPAR (peroxisome-proliferator-activated receptor)-gamma gene expression in brown adipocytes
|Lindgren EM,Nielsen R,Petrovic N,Jacobsson A,Mandrup S,Cannon B,Nedergaard J||The Biochemical journal (382:597)||2004|
Peroxisome proliferator-activated receptor alpha is downregulated in the failing human heart.
MA1-822 was used in western blot to investigate the role of peroxisome proliferator-activated receptor alpha in the failing human heart
|Karbowska J,Kochan Z,Smole¿ski RT||Cellular and molecular biology letters (8:49)||2003|
Effect of stages of lactation on the concentration of a 90-kilodalton heat shock protein in bovine mammary tissue.
MA1-822 was used in western blot to investigate the differences of HSP90 expression in cow mammary tissue at various lactation stages
|Watanabe A,Miyamoto T,Katoh N,Takahashi Y||Journal of dairy science (80:2372)||1997|
Peroxisome proliferator-activated receptor alpha physically interacts with CCAAT/enhancer binding protein (C/EBPbeta) to inhibit C/EBPbeta-responsive alpha1-acid glycoprotein gene expression.
MA1-822 was used in immunoprecipitation to study the interaction between peroxisome proliferator-activated receptor alpha and CCAAT/enhancer binding protein (C/EBPbeta) and its effect on alpha1 acid glycoprotein gene expression.
|Mouthiers A,Baillet A,Deloménie C,Porquet D,Mejdoubi-Charef N||Molecular endocrinology (Baltimore, Md.) (19:1135)||2005|
Evidence that peroxisome proliferator-activated receptor alpha is complexed with the 90-kDa heat shock protein and the hepatitis virus B X-associated protein 2.
MA1-822 was used in immunoprecipitation to investigate the interaction of PPARalpha with hsp90 and XAP2 and the function of XAP2.
|Sumanasekera WK,Tien ES,Turpey R,Vanden Heuvel JP,Perdew GH||The Journal of biological chemistry (278:4467)||2003|