Recombinant Human Thrombopoietin GMP Protein, CF GMP Best Seller

R&D Systems, part of Bio-Techne | Catalog # 288E-GMP

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Key Product Details

Source

E. coli

Accession #

NP_000451.1

Conjugate

Unconjugated

Applications

Bioactivity

View all Thrombopoietin/Tpo Proteins and Enzymes »

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).

Host Cell Protein

<5.00 ng per μg of protein when tested by ELISA.

Mycoplasma

Negative when tested in a ribosomal RNA hybridization assay.

Host Cell DNA

< 0.0010 ng per µg of protein when tested by PCR.

Scientific Data Images for Recombinant Human Thrombopoietin GMP Protein, CF

Recombinant Human Thrombopoietin GMP Protein Bioactivity

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 ED50 for 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

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

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 12 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.

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).

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. Foster, D. & Hunt, P. (1997) Thrombopoiesis and Thrombopoietins 13:203.
  16. Ehrenreich, H. et al. (2005) Proc. Natl. Acad. Sci. 102:862.
  17. 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

NP_000451.1

Additional Thrombopoietin/Tpo Products

Product Documents for Recombinant Human Thrombopoietin GMP Protein, CF

Product Datasheets

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.

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Manufacturing Specifications

R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Eu. Ph. 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products.


R&D Systems' quality focus includes:

  • Manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
  • Documented processes and QA control of documentation and process changes
  • Personnel training programs
  • Raw material testing and vendor qualification/monitoring
  • Fully validated equipment, processes and test methods
  • Equipment calibration schedules using a computerized calibration program
  • Facility maintenance, safety programs and pest control
  • Material review process for variances
  • Monitoring of stability over product shelf-life


R&D Systems strives to provide our customers with the analytical characteristics of each product so that customers may determine whether our products are appropriate for their research. The Certificate of Analysis provided contains the following lot specific information:

  • N-terminal amino acid analysis, SDS-PAGE analysis, and endotoxin level (as determined by LAL assay) performed on each bulk QC lot, not on individual bottlings of each QC lot
  • Post-bottling lot-specific bioassay results (compliance with an established range) and results of microbial testing according to USP
  • Host Cell Protein testing performed by ELISA
  • Mycoplasma testing by ribosomal RNA hybridization assay


Additional testing and documentation requested by the customer can be arranged at an additional cost.

Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis, Minnesota USA.

R&D Systems sells GMP grade products for preclinical or clinical ex vivo cell therapy applications. They are not for in vivo use. Please read the following End User Terms prior to using this product.

Animal-Free Manufacturing Conditions
Our dedicated controlled-access animal-free laboratories ensure that at no point in production are the products exposed to potential contamination by animal components or byproducts. Every stage of manufacturing is conducted in compliance with R&D Systems' stringent Standard Operating Procedures (SOPs). Production and purification procedures use equipment and media that are confirmed animal-free.

Production

  • All molecular biology procedures use animal-free media and dedicated labware.
  • Dedicated fermentors are utilized in committed animal-free areas.

Purification

  • Protein purification columns are animal-free.
  • Bulk proteins are filtered using animal-free filters.
  • Purified proteins are stored in animal-free containers in a dedicated cold storage room.

Quality Assurance

  • Low Endotoxin Level.
  • No impairment of biological activity.
  • High quality product obtained under stringent conditions.

Please read our complete Animal-Free Statement.

Product Specific Notices for Recombinant Human Thrombopoietin GMP Protein, CF

END USER TERMS OF USE OF PRODUCT

The following terms are offered to you upon your acceptance of these End User Terms of Use of Product. By using this product, you indicate your acknowledgment and agreement to these End User Terms of Use of Product. If you do not agree to be bound by and comply with all of the provisions of these End User Terms of Use of Product, you should contact your supplier of the product and make arrangements to return the product.

We suggest you print and retain a copy of these End User Terms of Use of Product for your records.

The End User is aware that R&D Systems, Inc. sells GMP products for preclinical or clinical ex vivo use and not for in vivo use. The End User further agrees, as a condition of the sale of R&D Systems' GMP products that: a) the End User will not use this GMP Product in any procedure wherein the product may be administered to humans, unless the End User has obtained, or prior to their use will have obtained, an Investigational New Drug (IND) exemption from the FDA and will use the product only in accordance with the protocols of such IND and of the Institutional Review Board overseeing the proposed research, or b) the End User will use the products outside of the United States in accordance with the protocols of research approved by the Institutional Review Board or authorized ethics committee and regulatory agencies to which the End User is subject to in their territory.

R&D Systems, Inc. has the right, at its sole discretion, to modify, add or remove any terms or conditions of these End User Terms of Use without notice or liability to you. Any changes to these End User Terms of Use are effective immediately following the printing of such changes on this product insert. The most recent version of these End User Terms of Use of Product may be found at: RnDSystems.com/Legal.

You agree to review these End User Terms of Use of Product to ensure any subsequent use by you of R&D Systems' GMP Products following changes to these End User Terms of Use of Product constitutes your acceptance of all such changes.

 

TERMS AND CONDITIONS

The following limitation applies to R&D Systems' warranty and liability for damages: All products are warranted to meet R&D Systems' published specifications when used under normal laboratory conditions.

R&D SYSTEMS DOES NOT MAKE ANY OTHER WARRANTY OR REPRESENTATION WHATSOEVER, WHETHER EXPRESS OR IMPLIED, WITH RESPECT TO ITS PRODUCTS. IN PARTICULAR, R&D SYSTEMS DOES NOT MAKE ANY WARRANTY OF SUITABILITY, NONINFRINGEMENT, MERCHANTABILITY OR FITNESS FOR ANY PARTICULAR PURPOSE.

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Full details of R&D Systems' Terms and Conditions of Sale can be found online at: RnDSystems.com/Legal.

For preclinical, or clinical ex vivo use

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