Immunofluorescence analysis of PPAR Gamma was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with PPAR gamma Rabbit Polyclonal Antibody (PA3821A) at 1ug/ml in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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 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||Pig, Rat, Mouse, Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues M(284) M G E D K I K F K H I T P L(298) of mouse PPAR gamma 2.|
|Storage buffer||whole serum|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunofluorescence (IF)||1-2 µg/ml|
|Western Blot (WB)||1:500-1:1500|
* 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.
PA3-821A detects peroxisome proliferator activated receptor (PPAR) gamma from human and mouse samples. This antibody is a reproduction of PA3-821, using the same immunizing peptide and new host rabbits.
PA3-821A has been successfully used in IF and Western blot procedures. Previous batches of this antibody have been used in gel shift, immunocytochemistry, and immunofluoresence procedures. By Western blot, this antibody detects a 57 kDa protein representing PPAR gamma 2 from differentiated mouse 3T3-L1 cell lysate. Immunolocalization of previous batches gives predominant nuclear staining.
The PA3-821A immunizing peptide corresponds to amino acid residues 284-298 from mouse PPAR gamma 2. This sequence is completely conserved in PPAR gamma 1, but exhibits no significant homology with PPAR alpha or NUC1. This sequence is completely conserved in mouse, human, and rat.
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. The PPAR gamma 2 isoform appears to be induced very early in the differentiation of several cultured adipocyte cell lines, and has been suggested to be a dominant regulator of the murine P2 (aP2) gene which encodes an intracellular lipid binding protein which is expressed only in adipose cells. PPAR gamma 2, like several other nuclear hormone receptors, heterodimerizes with RXR alpha.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Heterogeneous Differentiation of Human Mesenchymal Stem Cells in 3D Extracellular Matrix Composites.
PA3-821A was used in immunocytochemistry to analyze 3D extracellular matrix composites for heterogeneous differentiation of human mesenchymal stem cells
|Jung JP,Bache-Wiig MK,Provenzano PP,Ogle BM||BioResearch open access (5:37)||2016|
Human translocation liposarcoma-CCAAT/enhancer binding protein (C/EBP) homologous protein (TLS-CHOP) oncoprotein prevents adipocyte differentiation by directly interfering with C/EBPbeta function.
PA3-821 was used in immunocytochemistry to demonstrate that TLS-CHOP blocks adipocyte differentiation by directly interfering with C/EBPbeta function
|Adelmant G,Gilbert JD,Freytag SO||The Journal of biological chemistry (273:15574)||1998|
PPAR-¿ is repressed in Huntington's disease, is required for normal neuronal function and can be targeted therapeutically.
PA3-821A was used in immunohistochemistry and western blot to determine the role of decreased PPAR-delta in Huntington's disease and therapeutic targets
|Dickey AS,Pineda VV,Tsunemi T,Liu PP,Miranda HC,Gilmore-Hall SK,Lomas N,Sampat KR,Buttgereit A,Torres MJ,Flores AL,Arreola M,Arbez N,Akimov SS,Gaasterland T,Lazarowski ER,Ross CA,Yeo GW,Sopher BL,Magnuson GK,Pinkerton AB,Masliah E,La Spada AR||Nature medicine (22:37)||2016|
Urine acidification has no effect on peroxisome proliferator-activated receptor (PPAR) signaling or epidermal growth factor (EGF) expression in rat urinary bladder urothelium.
PA3-821A was used in immunohistochemistry to investigate the influence of urine acidification on signal pathways in urinary bladder urothelium
|Achanzar WE,Moyer CF,Marthaler LT,Gullo R,Chen SJ,French MH,Watson LM,Rhodes JW,Kozlosky JC,White MR,Foster WR,Burgun JJ,Car BD,Cosma GN,Dominick MA||Toxicology and applied pharmacology (223:246)||2007|
Dexamethasone induced preadipocyte recruitment and expression of CCAAT/enhancing binding protein alpha and peroxisome proliferator activated receptor-gamma proteins in porcine stromal-vascular (S-V) cell cultures obtained before and after the onset of fetal adipogenesis.
PA3-821A was used in immunohistochemistry to investigate the effect of dexamethasone on preadipocyte recruitment and transcription factor expression in stromal-vascular cells
|Hausman GJ||General and comparative endocrinology (133:61)||2003|
Thyroid hormone receptor sumoylation is required for preadipocyte differentiation and proliferation.
PA3-821A was used in ChIP assay to investigate thyroid hormone receptor in adipogenesis
|Liu YY,Ayers S,Milanesi A,Teng X,Rabi S,Akiba Y,Brent GA||The Journal of biological chemistry (290:7402)||2015|
Effects of lysophosphatidic acid on the in vitro proliferation and differentiation of a novel porcine preadipocyte cell line.
PA3-821A was used in western blot to investigate the influence of lysophosphatidic acid on pig preadipocyte cell proliferation and differentiation
|Nobusue H,Kondo D,Yamamoto M,Kano K||Comparative biochemistry and physiology. Part B, Biochemistry and molecular biology (157:401)||2010|
Activation of PPAR-¿ and PTEN cascade participates in lovastatin-mediated accelerated differentiation of oligodendrocyte progenitor cells.
PA3-821A was used in western blot to investigate the mechanism for the differentiation of oligodendrocyte progenitor cells induced by lovastatin
|Paintlia AS,Paintlia MK,Singh AK,Orak JK,Singh I||Glia (58:1669)||2010|
Ligand activation of peroxisome proliferator-activated receptor-beta/delta and inhibition of cyclooxygenase-2 enhances inhibition of skin tumorigenesis.
PA3-821A was used in western blot to investigate therapeutic benefit of nonsteroidal anti-inflammatory drugs on skin tumorigenesis and its molecular mechanism
|Bility MT,Zhu B,Kang BH,Gonzalez FJ,Peters JM||Toxicological sciences : an official journal of the Society of Toxicology (113:27)||2010|
Peroxisomal proliferator-activated receptor-gamma agonists induce partial reversion of epithelial-mesenchymal transition in anaplastic thyroid cancer cells.
PA3-821 was used in western blot to investigate the role of PPAR gamma in the tumorigenesis of anaplastic thyroid cancer cells.
|Aiello A,Pandini G,Frasca F,Conte E,Murabito A,Sacco A,Genua M,Vigneri R,Belfiore A||Endocrinology (147:4463)||2006|
Reciprocal expression of peroxisome proliferator-activated receptor-gamma and cyclooxygenase-2 in human term parturition.
PA3-821A was used in western blot to study the involvement of peroxisome proliferator-activated receptor-gamma and cyclooxygenase-2 in human term parturition
|Dunn-Albanese LR,Ackerman WE,Xie Y,Iams JD,Kniss DA||American journal of obstetrics and gynecology (190:809)||2004|
Enhanced growth inhibition by combination differentiation therapy with ligands of peroxisome proliferator-activated receptor-gamma and inhibitors of histone deacetylase in adenocarcinoma of the lung.
PA3-821 was used in western blot to investigate the role of the HDAC inhibitors in non-small cell lung cancer.
|Chang TH,Szabo E||Clinical cancer research : an official journal of the American Association for Cancer Research (8:1206)||2002|
MCF-7 and T47D human breast cancer cells contain a functional peroxisomal response.
PA3-821 was used in EMSA assay to characterize MCF-7 and T47D human breast cancer cells in terms of peroxisomal response
|Kilgore MW,Tate PL,Rai S,Sengoku E,Price TM||Molecular and cellular endocrinology (129:229)||1997|