Recombinant Human Thrombopoietin His (HEK-293) Protein, CF

Name: Recombinant Human Thrombopoietin His (HEK-293) Protein, CF
Brand: R&D Systems
SKU: 288-TPH

Catalog # 288-TPH | R&D Systems, Inc. a Bio-Techne Brand

Product Datasheet / COA / SDS

Analyzed by SEC-MALS. His-tag (HEK-293-expressed)

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Key Product Details

Source:	HEK293
Accession #:	P40225.1
Applications:	Bioactivity

View Full Technical Details for Recombinant Human Thrombopoietin His (HEK-293) Protein, CF

Technical Data
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Product Specifications for Recombinant Human Thrombopoietin His (HEK-293) Protein, CF

Source	Human embryonic kidney cell, HEK293-derived human Thrombopoietin/Tpo protein Ser22-Leu195, with a C-terminal 6-His tag
Purity	>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin level	<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Ser22
Predicted Molecular Mass	19 kDa
SDS-PAGE	19 kDa
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 0.0500-0.500 ng/mL.

Scientific Data Examples for Recombinant Human Thrombopoietin His (HEK-293) Protein, CF

	Recombinant Human Thrombopoietin/Tpo His-tag (HEK-293-expressed) Protein SEC-MALS. Recombinant Human Thrombopoietin (Catalog # 288-TPH) has a molecular weight (MW) of 23.3 kDa as analyzed by SEC-MALS, suggesting that this protein is a monomer. MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).
	Recombinant Human Thrombopoietin/Tpo His-tag (HEK-293-expressed) Protein Bioactivity. Recombinant Human Thrombopoietin/Tpo His-tag Protein (HEK-293-expressed) (Catalog # 288-TPH) stimulates proliferation in the MO7e human megakaryocytic leukemic cell line. The ED₅₀ for this effect is 0.0500‑0.500 ng/mL.
	Recombinant Human Thrombopoietin/Tpo His-tag (HEK-293-expressed) Protein SDS-PAGE. 2 μg/lane of Recombinant Human Thrombopoietin/Tpo His-tag (HEK-293-expressed) Protein (Catalog # 288-TPH) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 26-30 kDa.

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.

288-TPH

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute the 10 μg size at 100 μg/mL in PBS. Reconstitute all other sizes at 500 μg/mL in 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.

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The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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). 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).

Item	Value
References	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.
Entrez Gene IDs	7066 (Human); 21832 (Mouse); 81811 (Rat)
Alternate Names	MGDF, MGDFC-mpl ligand, MK-CSF, MKCSF, ML, MPL ligand, MPLLG, MPLLGMGC163194, Megakaryocyte colony-stimulating factor, Megakaryocyte growth and development factor, Myeloproliferative leukemia virus oncogene ligand, THCYT1, THPO, TPOMKCSF, Thrombopoietin, Tpo, megakaryocyte stimulating factor, thrombopoietin nirs variant 1

Citations for Recombinant Human Thrombopoietin His (HEK-293) Protein, CF

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Recombinant Human Thrombopoietin His (HEK-293) Protein, CF

Catalog # 288-TPH | R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

Product Specifications for Recombinant Human Thrombopoietin His (HEK-293) Protein, CF

Scientific Data Examples for Recombinant Human Thrombopoietin His (HEK-293) Protein, CF

Formulation, Preparation and Storage

Carrier Free

Reconstitution Calculator

Background: Thrombopoietin/Tpo

Citations for Recombinant Human Thrombopoietin His (HEK-293) Protein, CF

Product Documents for Recombinant Human Thrombopoietin His (HEK-293) Protein, CF

Product Datasheets

COA

Certificate of Analysis

SDS

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