Leptin, Quadruple Antagonist, 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, one polypeptide chain containing 146aa and additional Ala at N-terminus and having a molecular mass of ~16kD, oLEP was mutated, resulting in L39A/D40A/F41A/I42A mutant was purified by proprietary chromatographic techniques.\n\nAmino Acid Sequence: \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: \nThe purified ovine Leptin quadruple antagonist (16K) consists of > 95% monomers as determined by gel-filtration chromatography.\n\nBiological Activity:\nOvine LEP quadruple antagonist is capable of inhibiting leptin-induced proliferation of BAF/3 cells stably transfected with the long form of ovine leptin receptor. It also inhibits various leptin effects in several in vitro bioassays.\n\nEndotoxin: \nLess than 0.1ng/ug (IEU/ug) of ovine LEP triple antagonist\n\nProtein Content: \nProtein quantitation was carried out by UV spectroscopy at 280nm using the absorbency value of 0.21 as the extinction coefficient for a 0.1% (1mg/ml) solution at pH 8.0. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics).\n\nSolubility: \nIt is recommended to reconstitute the lyophilized LEP mutant in sterile 0.4% NaHCO3 adjusted to pH 8-9, 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 0.4% NaHCO3 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 after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

L1670-09A

Size

20ug

Form

Supplied as a sterile-filtered, lyophilized powder in 0.0045mM NaHCO3.

Purity

Purified by proprietary chromatographic techniques. (same/more than) 98.0% as determined by: (a) Gel filtration analysis. (b) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.

References

1. Chronic administration of recombinant ovine leptin in growing beef heifers: effects on secretion of LH, metabolic hormones, and timing of puberty. J Anim Sci 2004 Oct;82(10):2930-6.\n2. Overexpression of ovine leptin in Pichia pastoris: physiological yeast response to leptin production and characterization of the recombinant hormone. Yeast 2004 Feb;21(3):249-63.\n3. The late gestation increase in circulating ACTH and cortisol in the fetal sheep is suppressed by intracerebroventricular infusion of recombinant ovine leptin. J Endocrinol 2002 Aug;174(2):259-66.\n4. 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. Biol Reprod 2002 May;66(5):1555-61.\n5. 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. Domest Anim Endocrinol 2002 Apr;22(2):103-12.\n6. Effects of recombinant ovine leptin on in vitro lipolysis and lipogenesis in subcutaneous adipose tissue from lactating and nonlactating sheep. J Anim Sci 2001 Feb;79(2):445-52.