Human Thrombopoietin CHO Overexpression Lysate

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Human Thrombopoietin CHO Overexpression Lysate: 产品信息

产品描述
This Human Thrombopoietin overexpression lysate was created in CHO Cells and intented for use as a Western blot (WB) positive control. Purification of Thrombopoietin protein (Cat: 10381-H08C) from the overexpression lysate was verified.
表达宿主
CHO Cells
种属
Human
蛋白构建信息
A DNA sequence encoding the human THPO (NP_000451.1) precursor (Met 1-Gly 353) was fused with a polyhistidine tag at the C-terminus.
分子量
The recombinant human THPO consists of 343 amino acids with the predicted molecular mass of 36.9 kDa. As a result of glycosylation, the apparent molecular mass of rhTHPO is approximately 60-80 kDa in SDS-PAGE under reducing conditions.

Human Thrombopoietin CHO Overexpression Lysate: 使用指南

制备方法
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.
裂解缓冲液
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.
使用建议
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.
缓冲液
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
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.
应用
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human Thrombopoietin CHO Overexpression Lysate: 别称

Human MGDF Overexpression Lysate; Human MKCSF Overexpression Lysate; Human ML Overexpression Lysate; Human MPLLG Overexpression Lysate; Human THCYT1 Overexpression Lysate; Human TPO Overexpression Lysate

Thrombopoietin 背景信息

Thrombopoietin (TPO or THPO), also known as myeloproliferative leukemia virus ligand (c-Mpl), is a hematopoietic growth factor belonging to the EPO/TPO family. The thrombopoietin protein is produced mainly by the liver and the kidney that regulates the production of platelets by the bone marrow. Thrombopoietin protein stimulates both proliferation of progenitor megakaryocytes and their maturation to platelet-producing megakaryocytes, and also accelerates the recovery of platelets. Thrombopoietin protein is involved in cardiovascular disease as it regulates megakaryocyte development and enhances platelet adhesion/aggregation. It has been identified that surface c-MPL, the receptor for thrombopoietin protein, binds to the ligand and mediates the action.
全称
thrombopoietin
参考文献
  • Ryu T,et al.(2003) Thrombopoietin-producing hepatocellular carcinoma. Intern Med. 42(8): 730-4.
  • Higashihara M,et al. (2003) Thrombopoietin-producing tumor. Intern Med. 42(8): 632-3?
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