Recombinant Human Thrombopoietin, ACFP Protein

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ACFP288
R&D Systems Animal Component Free Recombinant Proteins
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Recombinant Human Thrombopoietin, ACFP Protein Summary

Product Specifications

Purity
>95%, 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.
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 0.3-3 ng/mL.
Source
Spodoptera frugiperda, Sf 9 (baculovirus)-derived human Thrombopoietin/Tpo protein
Ser22-Gly353
Produced in an animal component free process (ACFP).
Accession #
N-terminal Sequence
Analysis
Ser22
Predicted Molecular Mass
35 kDa
SDS-PAGE
40‑55 kDa, reducing conditions

Product Datasheets

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ACFP288 (carrier free)

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.

ACFP288

Formulation Lyophilized from a 0.2 μm filtered solution in HCl.
Reconstitution Reconstitute at 100 μg/mL in 4 mM HCl.
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.
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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.
Entrez Gene IDs
7066 (Human); 21832 (Mouse); 81811 (Rat)
Alternate Names
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

Manufacturing Specifications

Animal Component-Free Process (ACFP) Manufacturing Conditions
R&D Systems Animal Component-Free Process (ACFP) recombinant proteins are expressed in an animal-free certified Sf 9 insect cell line using dedicated animal-free raw materials and labware. Production and purification procedures use equipment and media that are confirmed animal-free but performed outside our dedicated animal-free laboratories. Every stage of the manufacturing process follows R&D Systems' stringent Standard Operating Procedures (SOPs). The certified Sf 9 insect cell bank has undergone extensive testing to certify the lack of cytopathogens by screening for various viruses, Mycoplasma, and Spiroplasmas using both in vitro and in vivo testing methods. For ex vivo research or bioproduction, additional documentation can be provided.

Please read our complete ACFP Statement
 

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