Resistin Human (E. coli), Tagless

Description

Total 93 AA. MW: 9.9 kDa (calculated). Three N-Terminal extra AA (highlighted).

Amino Acid Sequence

Source

E. coli

Purity

>95%

SDS-PAGE gel

12% SDS-PAGE separation of Human Resistin 1. M.W. marker – 14, 21, 31, 45, 66, 97 kDa 2. reduced and heated sample, 5μg/lane 3. non-reduced and non-heated sample, 5μg/lane

Formulation

Filtered (0,4 μm) and lyophilized in 0.5 mg/mL in 25mM Tris, 25mM NaCl, pH 7.5

Reconstitution

Add deionized water to prepare a working stock solution of approximately 0.5 mg/mL and let the lyophilized pellet dissolve completely. Product is not sterile! Please filter the product by an appropriate sterile filter before using it in the cell culture.

Storage, Stability/Shelf Life

Store lyophilized protein at –20°C. Lyophilized protein remains stable until the expiry date when stored at –20°C. Aliquot reconstituted protein to avoid repeated freezing/thawing cycles and store at –80°C for long term storage. Reconstituted protein can be stored at 4°C for a limited period of time; it does not show any change after two weeks at 4°C.

Quality Control Test

BCA to determine quantity of the protein.

SDS PAGE to determine purity of the protein.

Applications

ELISA, Western blotting

Introduction to the Molecule

Resistin, a product of the RSTN gene, is a peptide hormone belonging to the class of cysteine-rich secreted proteins which is termed the RELM family, and is also described as ADSF (Adipose Tissue-Specific Secretory Factor) and FIZZ3 (Found in Inflammatory Zone). Human resistin contains 108 amino acids as a prepeptide, and its hydrofobic signal peptide is cleaved before its secretion. Resistin circulates in human blood as a dimeric protein consisting of two 92 amino acid polypeptides, which are disulfide-linked via Cys26. Resistin may be an important link between obesity and insulin resistance. Mouse resistin, specifically produced and secreted by adipocyte, acts on skeletal muscle myocytes, hepatocytes and adipocytes themselves so that it reduces their sensitivity to insulin. Steppan et al. have suggested that resistin suppresses the ability of insulin to stimulace glucose uptake. They have also suggested that resistin is present at elevated levels in blood of obese mice, and is down regulated by fasting and antidiabetic drugs. Way et al., on the other hand, have found that resistin expression is severly suppressed in obesity and is stimulated by several antidiabetic drugs. Other studies have shown that mouse resistin increases during the differentiation of adipocytes, but it also seems to inhibit adipogenesis. In contrast, the human adipogenic differentiation is likely to be associated with a down regulation of resistin gene expression. Recent studies have shown that human resistin is expressed also in macrophages and may be a novel link between inflammation and insulin resistance.

References

• Kim KH. et al. A Cysteine-rich Adipose Tissue-specific Secretory Factors Inhibits Adipocte Differentiation. The Journal of Biological Chemistry. 276 (14), 11252-11256, (2001) .
• Steppan CM, Brown EJ, Wright CM, Bhat S, Banerjee RR, Dai CY, Enders GH, Silberg DG, Wen X, Wu GD, Lazar MA: . A Family of Tissue-specific Resistin-like Molecules. PNAS. 98, 502-506, (2001) .
• Steppan C.M. at al. A Family of Tissue-specific Resistin-like Molecules. Proc. Natl. Acad. Sci. USA. 98 (2), 502-506, (2001) .
• Way J.M. et al. Adipose Tissue Resistin Expression Is Severly Suppressed in Obesity and Stimulated by Peroxisome Proliferator-activated Receptor g Agonists. The Journal of Biological Chemistry. 276 (28), 25651-25653, (2001) .
• Way JM, Görgün CZ, Tong Q, Uysal KT, Brown KK, Harrington WW, Oliver WR Jr., Willson TM, Kliewer SA, Hotamisligil GS: . Adipose Tissue Resistin Expression Is Severly Suppressed in Obesity and Stimulated by Peroxisome Proliferator-activated Receptor γ Agonists. J Biol Chem. 276, 25651-25653, (2001) .
• Banerjee RR, Lazar MA: . Dimerization of resistin and resistin–like moleules is etermined by a single cystein. J Biol Chem. 276, 25970-25973, (2001) .
• Banerjee R.R., Lazar M.A. Dimerization of resistin and resistin–like moleules is etermined by a single cystein. The Journal of Biological Chemistry. 276, 25970-25973, (2001) .
• Maebuchi M, Machidori M, Urade R, Ogawa T, Moriyama T: . Low resistin levels in adipose tissues and serum high-fat fed mice and genetically obese mice: development of an ELISA system for quantification of resistin. Archiv Biochem Biophys. 416, 164-170 (2003) .
• Hartman HB, Hu X, Tyler KX, Dalal CK, Lazar MA: . Mechanisms Regulating Adipocyte Expression of Resistin. J Biol Chem. 277, 19754-19761, (2002) .
• Hartman H.B., Lazar M.A. at al. Mechanisms Regulating Adipocyte Expression of Resistin. The Journal of Biological Chemistry. 277 (22), 19754-19761, (2002) .
• Steppan CM, Lazar MA: . Resistin and obesity-associated insulin resistance. TRENDS Endocrinol Metab. 13, 18-23, (2002) .
• Steppan C.M., Lazar M.A. Resistin and obesity-associated insulin resistance. TRENDS in Endocrinology and Metabolism. 13 (1), 18-23, (2002) .
• Juan CC. at al. Suppressed Gene Expression of Adipocyte Resistin in an Insulin-resistant Rat Model Probably by Elevated Free Fatty Acids. Biochemical and Biophysical Research Communications. 289, 1328-1333, (2001) .
• Steppan CM, Bailley ST, Brown EJ, Banerjee RR, Wright CM, Patel HR, Ahima RS, Lazar MA: . The Hormone Links Obesity to Diabetes. Nature. 409, 307-312, (2001) .
• Steppan C.M. et al. The Hormone Links Obesity to Diabetes. Nature. 409, 307-312, (2001) .
• Pravenec M, Kazdová L, Landa V, Zídek V, Mlejnek P, Jansa P, Wang J, Qi N, Kurtz TW: . Transgenic and recombinant resistin impair skeletal muscle glucose metabolism in the spontaneously hypertensive rat. J Biol Chem. 278, 45209-45215, (2001) .
• Fasshauer M. at al. Tumor Necrosis Factor a Is a Negative Regulator of Resistin Gene Expression and Secretion in 3T3-L1 Adipocytes. Biochemical and Biophysical Research Communications. 208, 1027-1031, (2001) .