Recombinant Human Thrombopoietin (NS0-expressed) Protein

Catalog # 288-TPN | R&D Systems, Inc. a Bio-Techne Brand
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288-TPN-010/CF
288-TPN-025/CF
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288-TPN-100/CF
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288-TPN-025
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Key Product Details

Accession #

P40225

Source

NS0

Conjugate

Unconjugated

Applications

Bioactivity

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Thrombopoietin/Tpo protein
Ser22-Gly353

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ser22

Predicted Molecular Mass

35 kDa

SDS-PAGE

80-90 kDa, reducing conditions

Activity

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.

Scientific Data Images for Recombinant Human Thrombopoietin (NS0-expressed) Protein

Recombinant Human Thrombopoietin (NS0-expressed) Protein Bioactivity

Recombinant Human Thrombopoietin (NS0-expressed) Protein Bioactivity

Measured in a cell proliferation assay using MO7e human megakaryocytic leukemic cells. The ED50 for this effect is 5-15 ng/mL.

Formulation, Preparation and Storage

Carrier Free
What does CF mean?

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.

What formulation is right for me?

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.

Carrier: 288-TPN
Formulation Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Reconstitution Reconstitute at 50 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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 to -70 °C under sterile conditions after reconstitution.
Carrier Free: 288-TPN/CF
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 100 μg/mL in sterile PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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 to -70 °C under sterile conditions after reconstitution.

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). 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).

References

  1. Deutsch, V.R. and A. Tomer (2006) Br. J. Haematol. 134:453.
  2. Kaushansky, K. (2005) J. Clin. Invest. 115:3339.
  3. Li, J. et al. (1999) Br. J. Haematol. 106:345.
  4. Bartley, T.D. et al. (1994) Cell 77:1117.
  5. de Sauvage, F.J. et al. (1994) Nature 369:533.
  6. Marcucci, R. and M. Romano (2008) Biochim. Biophys. Acta 1782:427.
  7. Kaushansky, K. et al. (1994) Nature 369:568.
  8. Kaushansky, K. et al. (1995) Proc. Natl. Acad. Sci. 92:3234.
  9. Broudy, V.C. et al. (1995) Blood 85:1719.
  10. Lok, S.I. et al. (1994) Nature 369:565.
  11. Chen, J. et al. (1995) Blood 86:4054.
  12. Oda, A. et al. (1996) Blood 87:4664.
  13. Van Os, E. et al. (2003) Br. J. Haematol. 121:482.
  14. Kato, T. et al. (1997) Proc. Natl. Acad. Sci. 94:4669.
  15. Ehrenreich, H. et al. (2005) Proc. Natl. Acad. Sci. 102:862.
  16. Samoylenko, A. et al. (2008) Cell. Signal. 20:154.

Alternate Names

MGDF, MK-CSF, MKCSF, MPLLG, THCYT1, THPO, Tpo

Entrez Gene IDs

7066 (Human); 21832 (Mouse); 81811 (Rat)

Gene Symbol

THPO

UniProt

P40225

Product Documents for Recombinant Human Thrombopoietin (NS0-expressed) Protein

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS 288-TPN (with carrier)
Download SDS 288-TPN/CF (carrier free)

Product Specific Notices for Recombinant Human Thrombopoietin (NS0-expressed) Protein

For research use only

Citations for Recombinant Human Thrombopoietin (NS0-expressed) Protein (12)

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FAQs for Recombinant Human Thrombopoietin (NS0-expressed) Protein

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  • Q: Why do you sell Thrombopoietin protein from two different expression systems?  Under what circumstances would it be preferred to use the NS0-expressed protein?

    A: The NS0-expressed protein is necessary for the serum-free megakaryocyte colony stimulating assay (Nadine Weich, Genetics Insititute).  In all other assays, it works the same as the Sf-21 derived protein.

  • Q: What are the differences between the Recombinant Human Thrombopoietin/Tpo Proteins (Catalog #s 288-TP, 288-TPN, and 288-TPE)?

    A: 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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