Human SerpinD1 HEK293 Overexpression Lysate

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Human SerpinD1 HEK293 Overexpression Lysate: 製品の情報

製品の説明
This Human SerpinD1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of SerpinD1 protein (Cat: 10295-H08H) from the overexpression lysate was verified.
発現ホスト
HEK293 Cells
Human
タンパク質構築情報
A DNA sequence encoding the human SerpinD1 precursor (NP_000176.2) (Met 1-Ser 499) was expressed with a C-terminal polyhistidine tag.
分子量
The mature recombinant human SerpinD1 consists of 491 amino acids and predicts a molecular mass of 56.4 kDa. By SDS-PAGE under reducing conditions, the apparent molecular mass of rhSerpinD1 is approximately 65-70 kDa as a result of glycosylation.

Human SerpinD1 HEK293 Overexpression Lysate: 用法

調製方法
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
溶解バッファー
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
おすすめの用法
1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
バッファー
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
安定性 & 保存条件
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
アプリケーション
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human SerpinD1 HEK293 Overexpression Lysate: 別名

Human D22S673 Overexpression Lysate; Human HC2 Overexpression Lysate; Human HCF2 Overexpression Lysate; Human HCII Overexpression Lysate; Human HLS2 Overexpression Lysate; Human LS2 Overexpression Lysate; Human SerpinD1 Overexpression Lysate; Human THPH10 Overexpression Lysate

SerpinD1 背景情報

SerpinD1, also known as heparin cofactor II, is a member of Serpin superfamily of the serine proteinase inhibitors. HCII is a glycoprotein in human plasma that inhibits thrombin and chymotrypsin, and the rate of inhibition of thrombin is rapidly increased by Dermatan sulfate (DS), heparin (H) and glycosaminoglycans(GAG). The stimulatory effect of glycosaminoglycans on the inhibition is mediated, in part, by the N-terminal acidic domain of HCII. Interestingly, a C-terminal His-tagged recombinant HCII exhibits enhanced activity of thrombin inhibition. It has been suggested that HCII plays an unique and important role in vascular homeostasis, and accordingly mutations in this gene or congenital HCII deficiency is potentially associated with thrombosis. HCII specifically inhibits thrombin action at the site of vascular wall injury and HCII-thrombin complexes have been detected in human plasma. HCII protects against thrombin-induced vascular remodeling in both humans and mice and suggest that HCII is a predictive biomarker and therapeutic target for atherosclerosis. SerpinD1 also inhibits chymotrypsin, but in a glycosaminoglycan-independent manner.
完全な名称
serpin peptidase inhibitor, clade D (heparin cofactor), member 1
参考文献
  • Rau JC, et al. (2009) Heparin cofactor II in atherosclerotic lesions from the Pathobiological Determinants of Atherosclerosis in Youth (PDAY) study. Exp Mol Pathol. 87(3): 178-83.
  • Aihara K, et al. (2009) Heparin cofactor II as a novel vascular protective factor against atherosclerosis. J Atheroscler Thromb. 16(5): 523-31.
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