Leptin, Recombinant, Sheep (Ob, Obese Protein, Obesity Factor)

Leptin inhibits food intake and stimulates energy expenditure. Leptin also has thermogenic actions and regulates enzymes of fatty acid oxidation. Severe hereditary obesity in rodents and humans is caused by defects in leptin production. In addition to its critical role in the physiologic regulation of body weight leptin has a variety of other physiologic and pathologic functions resembling those of cytokines. These functions include the regulation of hematopoiesis, angiogenesis, wound healing, inflammation, and immune responses. \n\nRecombinant Ovine Leptin is a single, non-glycosylated, polypeptide chain containing 146 amino acids and having a molecular mass of 16,130 Dalton. \n\nSequence:\nThe sequence of the first five N-terminal amino acids was determined and was found to be Ala-Val-Pro-Ile-Arg. \n\nDimers and Aggregates:\n(same/less than) 1% as determined by silver-stained SDS-PAGE gel analysis. \n\nBiological Activity:\nLeptin is fully biologically active when compared to standards. The ED50, calculated by the leptin-dependent stimulation of Human OB-R transfected murine BaF3 indicator cells is 0.33-0.05ng/ml. \n\nEndotoxin:\n(same/less than) 0.1ng/ug (IEU/ug) of Leptin. \n\nProtein Content:\nProtein quantitation was carried out by two independent methods: \n1. UV spectroscopy at 280nm using the absorbency value of 0.2 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics). \n2. Analysis by RP-HPLC,using calibrated solution of Ovine Leptin as a Reference Standard. \n\nReconstitution:\nReconstitute the lyophilized Leptin in sterile 18M-cm H2O not less than 100ug/ml, which can then be further diluted to other aqueous solutions. \n\nStorage and Stability:\nLyophilized powder may be stored at 4 degrees C for short-term only. Reconstitute to nominal volume by adding sterile dH2O and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

L1670-16S

Size

100ug

Form

Supplied as a lyophilized powder in 0.0045mM NaHCO3.

Purity

(same/more than) 95% by RP-HPLC, FPLC, or reducing/non-reducing SDS-PAGE Silver Stain. Chromatographically purified.

References

1. Overexpression of ovine leptin in Pichia pastoris: physiological yeast response to leptin production and characterization of the recombinant hormone. Laborde C, Chemardin P, Combarnous Y, Yeast 2004 Feb;21(3):249-63 2. The late gestation increase in circulating ACTH and cortisol in the fetal sheep is suppressed by intracerebroventricular infusion of recombinant ovine leptin. Howe DC, Gertler A, J Endocrinol 2002 Aug;174(2):259-66 3. Central infusion of recombinant ovine leptin normalizes plasma insulin and stimulates a novel hypersecretion of luteinizing hormone after short-term fasting in mature beef cows. Amstalden M, Garcia MR, Nizielski SE, Biol Reprod 2002 May;66(5):1555-61 4. Effect of intravenous infusion of recombinant ovine leptin on feed intake and serum concentrations of GH, LH, insulin, IGF-1, cortisol, and thyroxine in growing prepubertal ewe lambs. Morrison CD, Wood R, Whitley NC, Domest Anim Endocrinol 2002 Apr;22(2):103-12 5. Effects of recombinant ovine leptin on in vitro lipolysis and lipogenesis in subcutaneous adipose tissue from lactating and nonlactating sheep. Newby D, Gertler A, J Anim Sci 2001 Feb;79(2):445-52 6. Adrenaline, insulin and glucagon do not have acute effects on plasma leptin levels in sheep: development and characterisation of an ovine leptin ELISA. Kauter K, Ball M, Tellam R, J Endocrinol 2000 Jul;166(1):127-35.