Adiponectin Polyclonal Antibody Catalog Number

Transcription

1 Website: thermofisher.com Customer Service (US): ext. 1 Technical Support (US): ext. 441 Adiponectin Polyclonal Antibody Catalog Number Details PA1-054 Size 200 µg Host/Isotope Class Type Immunogen Conjugate Form Concentration Purification Storage buffer Contains Storage Conditions Rabbit / IgG Polyclonal Antibody 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. Unconjugated Liquid 1 mg/ml Protein A PBS with 1mg/ml BSA 0.05% sodium azide -20 C, Avoid Freeze/Thaw Cycles Product data sheet Species Reactivity Tested species reactivity Mouse Published species reactivity Mouse, Human Tested Applications Dilution * Immunocytochemistry (ICC) 2 µg/ml Immunofluorescence (IF) 2 µg/ml Immunoprecipitation (IP) Assay dependent Western Blot (WB) 2 µg/ml Published Applications Western Blot (WB) See 13 publications below Immunohistochemistry (IHC) See 2 publications below Immunocytochemistry (ICC) See 1 publications below * 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. Product specific information 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. Background/Target Information 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.

2 Advanced Verification Data Adiponectin Antibody (PA1-054) in IAV Adiponectin Antibody (PA1-054) in TM Antibody specificity was demonstrated by showing that antibodies raised against the same target protein perform similarly. Western blot of Adiponectin using Anti-Adiponectin Polyclonal Antibody (Product # PA1-054), tested in parallel with Adiponectin ABfinity Oligoclonal Antibody (Product # ), shows similar expression of Adiponectin in the samples tested. Independent antibody validation info. Modulation of expression of target protein by cell treatment to demonstrate antibody specificity. Immunofluorescence analysis of Adiponectin using anti-adiponectin Polyclonal antibody (Product # PA1-054) shows increased expression of Leptin in differentiated 3T3-L1 cells. Cell Treatment validation info. Product Images For Adiponectin Polyclonal Antibody Adiponectin Antibody (PA1-054) in IF Immunofluorescence analysis of Adiponectin was performed using 70% confluent 3T3-L1 cells differentiated with StemPro Adipogenesis Supplement (Product # A ) 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 (Product # 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 (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 (Product # 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. Adiponectin Antibody (PA1-054) in WB Western blot analysis was performed on tissue extract (30 ug lysate) of Mouse Adipose (Lane 1). The blot was probed with Anti-Adiponectin Rabbit Polyclonal Antibody (Product # PA1-054, 2 ug/ml) and detected by chemiluminescence using Goat anti-rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.25 ug/ml, 1:4000 dilution). A 26 kda band corresponding to Adiponectin was observed in Mouse Adipose tissue. Known quantity of protein samples were electrophoresed using Novex NuPAGE 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002) and Novex Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane using the wet transfer system. The membrane was probed with the relevant primary and secondary antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

3 Adiponectin Antibody (PA1-054) in WB Western blot detection of Adiponectin from mouse serum using Product # PA1-054.

4 PubMed References For Adiponectin Polyclonal Antibody 13 Western Blot References PA1-054 was used in western blot to elucidate the relationship among FTO, IRX3 and fat metabolism. Mouse / 1:2500 Mouse / 1:2500 Scientific reports (Mar 2015; 5: null) "Fat mass- and obesity-associated gene Fto affects the dietary response in mouse white adipose tissue." Author(s):Ronkainen J,Huusko TJ,Soininen R,Mondini E,Cinti F,Mäkelä KA,Kovalainen M,Herzig KH,Järvelin MR,Sebert S, Savolainen MJ,Salonurmi T PubMed Article URL: 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 Stem cells (Dayton, Ohio) (Nov 2013; 31: 2528) "Mechanically activated Fyn utilizes mtorc2 to regulate RhoA and adipogenesis in mesenchymal stem cells." Author(s):Thompson WR,Guilluy C,Xie Z,Sen B,Brobst KE,Yen SS,Uzer G,Styner M,Case N,Burridge K,Rubin J PubMed Article URL: PA1-054 was used in western blot to study the modulation of adiponectin levels and oligimerization by lysyl hydroxylase-3 PloS one (Dec 2012; 7: null) "The activities of lysyl hydroxylase 3 (LH3) regulate the amount and oligomerization status of adiponectin." Author(s):Ruotsalainen H,Risteli M,Wang C,Wang Y,Karppinen M,Bergmann U,Kvist AP,Pospiech H,Herzig KH,Myllylä R PubMed Article URL: 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 Cardiovascular diabetology (Nov 2012; 11: null) "Telmisartan ameliorates insulin sensitivity by activating the AMPK/SIRT1 pathway in skeletal muscle of obese db/db mice." Author(s):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 PubMed Article URL: PA1-054 was used in western blot to study the activation of the AMPK-Sirt1 pathway in white adipose tissue by telmisartan European journal of pharmacology (Oct 2012; 692: 84) "Activation of AMPK-Sirt1 pathway by telmisartan in white adipose tissue: A possible link to anti-metabolic effects." Author(s):Shiota A,Shimabukuro M,Fukuda D,Soeki T,Sato H,Uematsu E,Hirata Y,Kurobe H,Sakaue H,Nakaya Y,Masuzaki H, Sata M PubMed Article URL: 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 Biochimica et biophysica acta (Apr 2012; 1822: 557) "St. John's Wort inhibits insulin signaling in murine and human adipocytes." Author(s):Richard AJ,Amini ZJ,Ribnicky DM,Stephens JM PubMed Article URL: PA1-054 was used in western blot to investigate the effect of STAT5A on adipogenesis Obesity (Silver Spring, Md.) (Sep 2011; 19: 1731) "STAT5A expression in Swiss 3T3 cells promotes adipogenesis in vivo in an athymic mice model system." Author(s):Stewart WC,Pearcy LA,Floyd ZE,Stephens JM PubMed Article URL: PA1-054 was used in western blot to characterize mouse adiponectin in terms of post-translational modification Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme (Jul 2011; 43: 545) "N-linked glycosylation of mouse adiponectin." Author(s):Tanaka M,Fukuhara A,Shimomura I PubMed Article URL:

5 PA1-054 was used in western blot to investigate the effect of glycosylation on adipogenesis Mouse / 1:1,000 Biochemical and biophysical research communications (Jul 2010; 398: 489) "Characteristic increase in nucleocytoplasmic protein glycosylation by O-GlcNAc in 3T3-L1 adipocyte differentiation." Author(s):Ishihara K,Takahashi I,Tsuchiya Y,Hasegawa M,Kamemura K PubMed Article URL: PA1-054 was used in western blot to study the role of Tribbles homologue 3 TRB3 during adipogenesis. Molecular and cellular biology (Oct 2007; 27: 6818) "TRB3 blocks adipocyte differentiation through the inhibition of C/EBPbeta transcriptional activity." Author(s):Bezy O,Vernochet C,Gesta S,Farmer SR,Kahn CR PubMed Article URL: PA1-054 was used in western blot to study the effect of rimonabant on endocrine function of mouse 3T3 F442A preadipocytes Molecular pharmacology (Feb 2006; 69: 471) "The cannabinoid CB1 receptor antagonist rimonabant (SR141716) inhibits cell proliferation and increases markers of adipocyte maturation in cultured mouse 3T3 F442A preadipocytes." Author(s):Gary-Bobo M,Elachouri G,Scatton B,Le Fur G,Oury-Donat F,Bensaid M PubMed Article URL: 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. The Journal of biological chemistry (Oct 2005; 280: 35361) "JNK and tumor necrosis factor-alpha mediate free fatty acid-induced insulin resistance in 3T3-L1 adipocytes." Author(s):Nguyen MT,Satoh H,Favelyukis S,Babendure JL,Imamura T,Sbodio JI,Zalevsky J,Dahiyat BI,Chi NW,Olefsky JM PubMed Article URL: PA1-054 was used in western blot to investigate the function of EHD1 and EHBP1 in perinuclear localization of GLUT4. 2 Immunohistochemistry References The Journal of biological chemistry (Sep 2004; 279: 40062) "Role of EHD1 and EHBP1 in perinuclear sorting and insulin-regulated GLUT4 recycling in 3T3-L1 adipocytes." Author(s):Guilherme A,Soriano NA,Furcinitti PS,Czech MP PubMed Article URL: PA1-054 was used in immunohistochemistry to study the role of T-cadherin in the mechanism by which adiponectin promotes revascularization The Journal of biological chemistry (Aug 2013; 288: 24886) "T-cadherin is essential for adiponectin-mediated revascularization." Author(s):Parker-Duffen JL,Nakamura K,Silver M,Kikuchi R,Tigges U,Yoshida S,Denzel MS,Ranscht B,Walsh K PubMed Article URL: PA1-054 was used in immunohistochemistry to study the role of T-cadherin in angiogenesis. 1 Immunocytochemistry References Cancer research (Mar 2008; 68: 1407) "T-cadherin supports angiogenesis and adiponectin association with the vasculature in a mouse mammary tumor model." Author(s):Hebbard LW,Garlatti M,Young LJ,Cardiff RD,Oshima RG,Ranscht B PubMed Article URL: PA1-054 was used in immunocytochemistry to investigate the effect of adiponectin on tumor angiogenesis in in a mammary cancer mouse model. Human / Not Cited Clinical cancer research : an official journal of the American Association for Cancer Research (May 2009; 15: 3256) "Adiponectin deficiency limits tumor vascularization in the MMTV-PyV-mT mouse model of mammary cancer." Author(s):Denzel MS,Hebbard LW,Shostak G,Shapiro L,Cardiff RD,Ranscht B PubMed Article URL: