Human Thrombopoietin HEK293 Overexpression Lysate: Product Information
This Human Thrombopoietin overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Thrombopoietin protein (Cat: 10381-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human THPO (NP_000451.1) (Met1-Gly353) was expressed with a polyhistidine tag at the C-terminus.
The recombinant human THPO consists 182 amino acids and predicts a molecular mass of 19.8 kDa.
Human Thrombopoietin HEK293 Overexpression Lysate: Usage Guide
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.
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.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
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.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human Thrombopoietin HEK293 Overexpression Lysate: Alternative Names
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 Background Information
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.
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