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GDF-8, MSTN, Growth/Differentiation Factor 8,MSTN Muscle Hypertrophy.
Myostatin (GDF-8) is expressed uniquely in human skeletal muscle as a 12 kDa mature glycoprotein consisting of 113 amino acid residues and secreted into plasma. Myostatin is a member of the transforming growth factor β superfamily of secreted growth and differentiation factors that is essential for proper regulation of skeletal muscle mass. Studies have shown that myostatin could play an important role in cardiac development and physiology.
Total 152AA. M.W. 16kDa (calculated). N-terminal His-tag and spacer (39AA – underlined) MRGSHHHHHH GMASMTGGQQ MGRDLYDDDD KDPSSRSAXR SRRDFGLDCD EHSTESRCCR YPLTVDFEAF GWDWIIAPKR YKANYCSGEC EFVFLQKYPH THLVHQANPR GSAGPCCTPT KMSPINMLYF NGKEQIIYGK IPAMVVDRCG CS.
Escherichia Coli.
The human myostatin part of the fusion protein is corresponding to amino acidsequence of the human myostatin Swiss-Prot- O14793, R263‑S375).
Three-step procedure using affinity Ni-NTA chromatography and size exclusion chromatography before and after refolding
Sterile filtered and lyophilized from 0.5 mg/ml in 0.05 M acetate buffer, pH 5.5.
Add 0.2 ml of 0.1 M acetate buffer pH 4 and let the lyophilized pellet dissolve completely. The lyophilized myostatin is poorly soluble at neutral pH.
Greater than 95% (SDS-PAGE analyzed).
Store lyophilized protein at -20°C. Aliquot the product after reconstitution to avoid repeated freezing/thawing cycles. 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.
The lyophilized GDF-8 remains stable until the expiry date when stored at -20°C.
Western blotting, ELISA.
• Artaza JN, Bhasin S, Magee TR, Reisz-Porszasz S, Shen R, Groome NP, Fareez MM, Gonzalez-Cadavid NF. Myostatin inhibits myogenesis and promotes adipogenesis in C3H 10T(1/2) mesenchymal multipotent cells. Endocrinology. 2005 Aug;146(8): 3547-57. Epub 2005 May 5.
• Taylor WE, Bhasin S, Artaza J, Byhower F, Azam M, Willard DH Jr, Kull FC Jr, Gonzalez-Cadavid N. Myostatin inhibits cell proliferation and protein synthesis in C2C12 muscle cells. Am J Physiol Endocrinol Metab. 2001 Feb; 280(2):E221-8.
• McPherron AC, Lee SJ. Double muscling in cattle due to mutations in the myostatin gene. Proc Natl Acad Sci U S A. 1997 Nov 11;94(23):12457-61.
• Sharma M, Kambadur R, Matthews KG, Somers WG, Devlin GP, Conaglen JV, Fowke PJ, Bass JJ. Myostatin, a transforming growth factor-beta superfamily member, is expressed in heart muscle and is upregulated in cardiomyocytes after infarct. J Cell Physiol. 1999 Jul;180(1):1-9.
• Gonzalez-Cadavid NF, Taylor WE, Yarasheski K, Sinha-Hikim I, Ma K, Ezzat S, Shen R, Lalani R, Asa S, Mamita M, Nair G, Arver S, Bhasin S. Organization of the human myostatin gene and expression in healthy men and HIV-infected men with muscle wasting. Proc Natl Acad Sci U S A. 1998 Dec 8;95(25):14938-43.
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