Recombinant Human Thrombopoietin GMP Protein, CFGMP

Name: Recombinant Human Thrombopoietin GMP Protein, CF
Brand: rnd
SKU: 288E-GMP

Catalog # 288E-GMP

Datasheet / COA / SDS

Animal Free

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Key Product Details

Source:	E. coli
Accession #:	NP_000451
Applications:	Bioactivity

View Full Technical Details for Recombinant Human Thrombopoietin GMP Protein, CF

Technical Data
Citations / Reviews / FAQs

Product Specifications

Source	E. coli-derived human Thrombopoietin/Tpo protein Ser22-Leu195 Produced in an animal-free laboratory. Manufactured and tested under cGMP guidelines
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	Ala-Ser22-Pro-Ala-Pro-Pro-Ala-(Cys)-Asp-Leu
Predicted Molecular Mass	18.7 kDa
SDS-PAGE	19 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 0.05-0.5 ng/mL. The specific activity of Recombinant Human Thrombopoietin is >1 x 10⁷ units/mg, which is calibrated against the human Thrombopoietin reference standard (NIBSC code: 03/124).

Recombinant Human Thrombopoietin GMP Protein, CF Scientific Data Examples

	Recombinant Human Thrombopoietin GMP Protein Bioactivity GMP-grade Recombinant Human Thrombopoietin (Catalog # 288E-GMP) stimulates proliferation in the MO7e human megakaryocytic leukemic cell line. The ED₅₀for this effect is 0.05-0.5 ng/mL. Three independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of GMP Thrombopoietin.
	Recombinant Human Thrombopoietin GMP Protein SDS-PAGE 2 μg/lane of GMP-grade Recombinant Human Thrombopoietin (Catalog # 288E-GMP) 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.

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.

288E-GMP

Formulation	Lyophilized from a 0.2 μm filtered solution in Sodium Acetate.
Reconstitution	Reconstitute at 100-200 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. A minimum of 6 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, ≤ -20 °C under sterile conditions after reconstitution.

Reconstitution Calculator

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.< /li>
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

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