Immunofluorescence analysis of Adiponectin was performed using 70% confluent 3T3-L1 cells differentiated with StemPro Adipogenesis Supplement (A10065-01) for 5 days. 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 Adiponectin Rabbit Polyclonal Antibody (PA1-054) at 2 µg/ml in 0.1% 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 conj µgate (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 represents the merged image showing cytoplasmic localization. Panel e is untreated cell with no signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
|Tested species reactivity||Mouse|
|Published species reactivity||Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptides corresponding to the residues E(18) D D V T T T E E L A P A L V(32) and F(187) T Y D Q Y Q E K N V D Q A(200) of mouse Adiponectin.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||4-8 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.
PA1-054 detects Adiponectin (30 kDa) from mouse serum.
PA1-054 has been successfully used in Western blot and immunoprecipitation procedures. By Western blot, this antibody detects an ~30 kDa protein representing Adiponectin from mouse serum.
The PA1-054 immunogen is a pair of synthetic peptides corresponding to the residues E(18) D D V T T T E E L A P A L V(32) and F(187) T Y D Q Y Q E K N V D Q A(200) of mouse Adiponectin.
Adipose cells produce and secrete numerous physiologically important proteins, such as lipoprotein lipase (LPL), leptin and adiponectin protein of 30 kDa. Adiponectin is a circulating protein that is secreted exclusively by differentiated adipocytes. During adipocyte differentiation, mRNA levels of Adiponectin have been shown to be induced over 100-fold. Studies indicate that Adiponectin enhances the ability of sub-physiological levels of insulin to suppress glucose production, thus linking adipose tissue to whole body glucose regulation. Adiponectin function appears to be regulated by phosphatidylinositol-3-kinase (PI3-K) since Adiponectin secretion is blocked by pharmacologic inhibitors of this kinase. mRNA levels of Adiponectin have been shown to be significantly reduced in adipose tissue of obese patients with Type 2 diabetes relative to control subjects. Structural similarities to TNFalpha suggests that Adiponectin may play a role in pathogenesis of insulin resistance in Type 2 diabetes. Because Adiponectin is down-regulated in various forms of obesity and its structural similarity to TNFalpha, it is currently being investigated as an important regulator of whole body energy homeostasis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Fat mass- and obesity-associated gene Fto affects the dietary response in mouse white adipose tissue.
PA1-054 was used in western blot to elucidate the relationship among FTO, IRX3 and fat metabolism.
|Ronkainen J,Huusko TJ,Soininen R,Mondini E,Cinti F,Mäkelä KA,Kovalainen M,Herzig KH,Järvelin MR,Sebert S,Savolainen MJ,Salonurmi T||Scientific reports (5:null)||2015|
Mechanically activated Fyn utilizes mTORC2 to regulate RhoA and adipogenesis in mesenchymal stem cells.
PA1-054 was used in western blot to study the regulation of adipogenesis by mechanical strain in mesenchymal stem cells and the roles played by Fyn, mTORC2 and RhoA
|Thompson WR,Guilluy C,Xie Z,Sen B,Brobst KE,Yen SS,Uzer G,Styner M,Case N,Burridge K,Rubin J||Stem cells (Dayton, Ohio) (31:2528)||2013|
The activities of lysyl hydroxylase 3 (LH3) regulate the amount and oligomerization status of adiponectin.
PA1-054 was used in western blot to study the modulation of adiponectin levels and oligimerization by lysyl hydroxylase-3
|Ruotsalainen H,Risteli M,Wang C,Wang Y,Karppinen M,Bergmann U,Kvist AP,Pospiech H,Herzig KH,Myllylä R||PloS one (7:null)||2012|
Telmisartan ameliorates insulin sensitivity by activating the AMPK/SIRT1 pathway in skeletal muscle of obese db/db mice.
PA1-054 was used in western blot to study the role of the AMPK/SIRT1 pathway in the mechanism by which telmisartan improves skeletal muscle insulin-sensitivity in obese mice
|Shiota A,Shimabukuro M,Fukuda D,Soeki T,Sato H,Uematsu E,Hirata Y,Kurobe H,Maeda N,Sakaue H,Masuzaki H,Shimomura I,Sata M||Cardiovascular diabetology (11:null)||2012|
Activation of AMPK-Sirt1 pathway by telmisartan in white adipose tissue: A possible link to anti-metabolic effects.
PA1-054 was used in western blot to study the activation of the AMPK-Sirt1 pathway in white adipose tissue by telmisartan
|Shiota A,Shimabukuro M,Fukuda D,Soeki T,Sato H,Uematsu E,Hirata Y,Kurobe H,Sakaue H,Nakaya Y,Masuzaki H,Sata M||European journal of pharmacology (692:84)||2012|
St. John's Wort inhibits insulin signaling in murine and human adipocytes.
PA1-054 was used in western blot to study the inhibition of human and murine adipocyte insulin signaling by extracts of St John's Wort
|Richard AJ,Amini ZJ,Ribnicky DM,Stephens JM||Biochimica et biophysica acta (1822:557)||2012|
STAT5A expression in Swiss 3T3 cells promotes adipogenesis in vivo in an athymic mice model system.
PA1-054 was used in western blot to investigate the effect of STAT5A on adipogenesis
|Stewart WC,Pearcy LA,Floyd ZE,Stephens JM||Obesity (Silver Spring, Md.) (19:1731)||2011|
N-linked glycosylation of mouse adiponectin.
PA1-054 was used in western blot to characterize mouse adiponectin in terms of post-translational modification
|Tanaka M,Fukuhara A,Shimomura I||Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme (43:545)||2011|
Characteristic increase in nucleocytoplasmic protein glycosylation by O-GlcNAc in 3T3-L1 adipocyte differentiation.
PA1-054 was used in western blot to investigate the effect of glycosylation on adipogenesis
|Ishihara K,Takahashi I,Tsuchiya Y,Hasegawa M,Kamemura K||Biochemical and biophysical research communications (398:489)||2010|
TRB3 blocks adipocyte differentiation through the inhibition of C/EBPbeta transcriptional activity.
PA1-054 was used in western blot to study the role of Tribbles homologue 3 TRB3 during adipogenesis.
|Bezy O,Vernochet C,Gesta S,Farmer SR,Kahn CR||Molecular and cellular biology (27:6818)||2007|
The cannabinoid CB1 receptor antagonist rimonabant (SR141716) inhibits cell proliferation and increases markers of adipocyte maturation in cultured mouse 3T3 F442A preadipocytes.
PA1-054 was used in western blot to study the effect of rimonabant on endocrine function of mouse 3T3 F442A preadipocytes
|Gary-Bobo M,Elachouri G,Scatton B,Le Fur G,Oury-Donat F,Bensaid M||Molecular pharmacology (69:471)||2006|
JNK and tumor necrosis factor-alpha mediate free fatty acid-induced insulin resistance in 3T3-L1 adipocytes.
PA1-054 was used in western blot to investigate the role of JNK and tumor necrosis factor-alpha in free fatty acid-induced insulin resistance in 3T3-L1 adipocytes.
|Nguyen MT,Satoh H,Favelyukis S,Babendure JL,Imamura T,Sbodio JI,Zalevsky J,Dahiyat BI,Chi NW,Olefsky JM||The Journal of biological chemistry (280:35361)||2005|
Role of EHD1 and EHBP1 in perinuclear sorting and insulin-regulated GLUT4 recycling in 3T3-L1 adipocytes.
PA1-054 was used in western blot to investigate the function of EHD1 and EHBP1 in perinuclear localization of GLUT4.
|Guilherme A,Soriano NA,Furcinitti PS,Czech MP||The Journal of biological chemistry (279:40062)||2004|
T-cadherin is essential for adiponectin-mediated revascularization.
PA1-054 was used in immunohistochemistry to study the role of T-cadherin in the mechanism by which adiponectin promotes revascularization
|Parker-Duffen JL,Nakamura K,Silver M,Kikuchi R,Tigges U,Yoshida S,Denzel MS,Ranscht B,Walsh K||The Journal of biological chemistry (288:24886)||2013|
T-cadherin supports angiogenesis and adiponectin association with the vasculature in a mouse mammary tumor model.
PA1-054 was used in immunohistochemistry to study the role of T-cadherin in angiogenesis.
|Hebbard LW,Garlatti M,Young LJ,Cardiff RD,Oshima RG,Ranscht B||Cancer research (68:1407)||2008|
Adiponectin deficiency limits tumor vascularization in the MMTV-PyV-mT mouse model of mammary cancer.
PA1-054 was used in immunocytochemistry to investigate the effect of adiponectin on tumor angiogenesis in in a mammary cancer mouse model.
|Denzel MS,Hebbard LW,Shostak G,Shapiro L,Cardiff RD,Ranscht B||Clinical cancer research : an official journal of the American Association for Cancer Research (15:3256)||2009|