Recombinant Human Thrombopoietin (NS0-expressed) Protein Summary
Details of Functionality
Measured in a cell proliferation assay using MO7e human megakaryocytic leukemic cells. Avanzi, G. et al. (1988) Br. J. Haematol. 69:359. The ED50 for this effect is 5-15 ng/mL.
Source
Mouse myeloma cell line, NS0-derived human Thrombopoietin/Tpo protein Ser22-Gly353
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Note
<0.10 EU per 1 μg of the protein by the LAL method.
Applications/Dilutions
Dilutions
Bioactivity
Theoretical MW
35 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
80-90 kDa, reducing conditions
Publications
Read Publications using 288-TPN in the following applications:
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
3 months, -20 to -70 °C under sterile conditions after reconstitution.
Buffer
Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Reconstitution Instructions
Reconstitute at 50 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.
Notes
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Human Thrombopoietin (NS0-expressed) Protein
Megakaryocyte colony-stimulating factor
Megakaryocyte growth and development factor
megakaryocyte stimulating factor
MGDF
MGDFC-mpl ligand
MKCSF
MK-CSF
ML
MPL ligand
MPLLG
MPLLGMGC163194
Myeloproliferative leukemia virus oncogene ligand
THCYT1
THPO
thrombopoietin nirs variant 1
Thrombopoietin
Tpo
TPOMKCSF
Background
Thrombopoietin (Tpo), is a key regulator of megakaryocytopoiesis and thrombopoiesis. It is principally produced in the liver and is bound and internalized by the receptor Tpo R/c-mpl. Defects in the Tpo-Tpo R signaling pathway are associated with a variety of platelet disorders (1 - 3). The 353 amino acid (aa) human Tpo precursor is cleaved to yield the 332 aa mature protein. Mature human Tpo shares approximately 70% aa sequence homology with mouse and rat Tpo. It is an 80 - 85 kDa protein that consists of an N-terminal domain with homology to Erythropoietin (Epo) and a C-terminal domain that contains multiple N-linked and O-linked glycosylation sites (4, 5). Tissue specific alternate splicing of human Tpo generates multiple isoforms with internal deletions, insertions, and/or C-terminal substitutions (6). Tpo promotes the differentiation, proliferation, and maturation of MK and their progenitors (4, 5, 7). Several other cytokines can promote these functions as well but only in cooperation with Tpo (8, 9). Notably, IL-3 independently induces MK development, although its effects are restricted to early in the MK lineage (8, 9). Tpo additionally promotes platelet production, aggregation, ECM adhesion, and activation (10 - 13). It is cleaved by platelet-derived thrombin following Arg191 within the C-terminal domain and subsequently at other sites upon extended digestion (14). Full length Tpo and shorter forms circulate in the plasma (4, 5). The C terminal domain is not required for binding to Tpo R or inducing MK growth and differentiation (5). Aside from its hematopoietic effects, Tpo is expressed in the brain where it promotes the apoptosis of hypoxia-sensitized neurons and inhibits neuronal differentiation by blocking NGF induced signaling (15, 16).
Deutsch, V.R. and A. Tomer (2006) Br. J. Haematol. 134:453.
Kaushansky, K. (2005) J. Clin. Invest. 115:3339.
Li, J. et al. (1999) Br. J. Haematol. 106:345.
Bartley, T.D. et al. (1994) Cell 77:1117.
de Sauvage, F.J. et al. (1994) Nature 369:533.
Marcucci, R. and M. Romano (2008) Biochim. Biophys. Acta 1782:427.
Kaushansky, K. et al. (1994) Nature 369:568.
Kaushansky, K. et al. (1995) Proc. Natl. Acad. Sci. 92:3234.
Broudy, V.C. et al. (1995) Blood 85:1719.
Lok, S.I. et al. (1994) Nature 369:565.
Chen, J. et al. (1995) Blood 86:4054.
Oda, A. et al. (1996) Blood 87:4664.
Van Os, E. et al. (2003) Br. J. Haematol. 121:482.
Kato, T. et al. (1997) Proc. Natl. Acad. Sci. 94:4669.
Ehrenreich, H. et al. (2005) Proc. Natl. Acad. Sci. 102:862.
Samoylenko, A. et al. (2008) Cell. Signal. 20:154.
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