Recombinant Human Thrombopoietin (E. coli-expressed), CF
Recombinant Human Thrombopoietin (E. coli-expressed), CF Summary
Product Specifications
Ser22-Leu195
Analysis
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
288-TPE
| Formulation | Lyophilized from a 0.2 μm filtered solution in Sodium Acetate. |
| Reconstitution | Reconstitute at 100 μg/mL in sterile, deionized water. |
| Shipping | The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Scientific Data
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Recombinant Human Thrombopoietin/Tpo (Catalog # 288-TPE) stimulates proliferation in the MO7e human megakaryocytic leukemic cell line. The ED50 for this effect is 0.05-0.5 ng/mL, which is more than 2-fold more active than two competitors'.
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2 μg/lane of Recombinant Human Thrombopoietin/Tpo was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 19 kDa and 18 kDa, respectively.
Reconstitution Calculator
Background: Thrombopoietin/Tpo
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).
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- 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.
FAQs
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What are the differences between the Recombinant Human Thrombopoietin/Tpo Proteins (Catalog #s 288-TP, 288-TPN, and 288-TPE)?
Each of these proteins is produced using a different expression system. 288-TP is Spodoptera frugiperda, Sf21 baculovirus-derived; 288-TPN is mouse myeloma cell line, NS0-derived; and 288-TPE is E. coli-derived.
Additionally, the sequence length also varies. 288-TP and 288-TPN begin at Ser22 and end at Gly353. 288-TPE also begins at Ser22 but is truncated at the C-terminus, ending at Leu195. Truncation of the C-terminus has been shown to enhance the biological activity of Tpo.
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