Recombinant Human Thrombopoietin (E. coli-expressed), CF 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 0.05-0.5 ng/mL.
|
Source |
E. coli-derived human Thrombopoietin/Tpo protein Ser22-Leu195 |
Accession # |
|
N-terminal Sequence |
Ala-Ser22
|
Protein/Peptide Type |
Recombinant Proteins |
Purity |
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. |
Endotoxin Note |
<0.10 EU per 1 μg of the protein by the LAL method. |
Applications/Dilutions
Dilutions |
|
Theoretical MW |
18.7 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 |
19 kDa, reducing conditions
|
Packaging, Storage & Formulations
Storage |
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 °C under sterile conditions after reconstitution.
|
Buffer |
Lyophilized from a 0.2 μm filtered solution in Sodium Acetate. |
Purity |
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. |
Reconstitution Instructions |
Reconstitute at 100 μg/mL in sterile, deionized water. |
Notes
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Human Thrombopoietin (E. coli-expressed), CF
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).
Both full length Tpo and shorter forms circulate in the plasma, with the
shorter, N‑terminal EPO-like domain forms showing significantly increased
specific activity (4, 5, 15). 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 (16, 17).
- 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.
- Foster, D. & Hunt, P. (1997) Thrombopoiesis and Thrombopoietins 13:203.
- Ehrenreich, H. et al. (2005) Proc. Natl. Acad. Sci. 102:862.
- Samoylenko, A. et al. (2008) Cell. Signal. 20:154.
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