Recombinant Human Thrombopoietin/TPO GMP Protein, CF

The bioactivity of Recombinant Human Thrombopoietin/TPO GMP Protein (Catalog # BT-TPO-GMP) was measured in a cell proliferation assay using MO7e human megakaryocytic leukemic cells. The ED50 for this effect is 0.0500‑0.500 ng/mL. Three independent lots were tested for bioactivity and plotted on the same graph to show lot-to-lot consistency of GMP Thrombopoietin/Tpo protein.

Equivalent bioactivity of GMP (Catalog # BT-TPO-GMP) and Animal-Free (BT-TPO-AFL) grades of Recombinant Human Thrombopoietin/TPO as measured in a cell proliferation assay using MO7e human megakaryocytic leukemic cells (orange and green, respectively).

2 μg/lane of Recombinant Human Thrombopoietin/TPO GMP Protein (Catalog # BT-TPO-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 18-19 kDa under reducing conditions.

Purified CD34+ cells derived from plerixafor mobilized peripheral blood were cultured in commercially available media. The media was supplemented with SCF (100 ng/mL, Catalog # BT-SCF-GMP), TPO (100 ng/mL, Catalog # BT-TPO-GMP), FLT-3L (100 ng/mL, Catalog # BT-FT3L-GMP), and SR1 (1 µM, Catalog # 7086). After 7 days, fold expansion was calculated from total viable cell counts. Frequency (%) CD34+ cells was analyzed by flow cytometry on Days 0 and 7. The average fold expansion of n=2 technical replicates is shown.​

Purified CD34+ cells derived from plerixafor mobilized peripheral blood were cultured in commercially available media. The media was supplemented with SCF (Catalog # BT-SCF-GMP, 100 ng/mL), TPO (Catalog # BT-TPO-GMP, 100 ng/mL), FLT-3L (Catalog # BT-FT3L-GMP, 100 ng/mL), and SR1 (Catalog # 7086, 1 µM). After 7 days, fold expansion was calculated from total viable cell counts. Percent (%) CD34+ cells was analyzed by flow cytometry on Days 0 and 7. The average fold expansion of n=3 technical replicates is shown.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human Thrombopoietin/TPO GMP Protein, CF Summary

• Details of Functionality: 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.
  The specific activity of Recombinant Human Thrombopoietin is >1.00 x 10⁷ units/mg, which is calibrated against the Human Thrombopoietin reference standard (NIBSC code: 03/124).
• Source: E. coli-derived human Thrombopoietin/Tpo protein
  Ser22 - Leu195 with an N-terminal Alanine
  Produced using non-animal reagents in an animal-free laboratory.
  Manufactured and tested under cGMP guidelines.
• Accession #: NP_000451.1
• N-terminal Sequence: Ala-Ser22-Pro-Ala-Pro-Pro-Ala-(Cys)-Asp-Leu
• Protein/Peptide Type: GMP Recombinant Proteins
• Purity: >95%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. 18732 Da ± 50 Da
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 18.7 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 18-19 kDa, under reducing conditions.

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in Acetonitrile and TFA with Trehalose.
• Purity: >95%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. 18732 Da ± 50 Da
• Reconstitution Instructions: Reconstitute at 500 μg/mL in sterile water.

Notes

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Alternate Names for Recombinant Human Thrombopoietin/TPO GMP Protein, CF

• 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

Background

Thrombopoietin (TPO) is a crucial regulator of hematopoietic stem cell (HSC) differentiation, maturation, and proliferation, as well as megakaryocytopoiesis and thrombopoiesis. TPO is often used in conjunction with Stem Cell Factor (SCF) and Flt-3 ligand in cell culture protocols to expand HSCs for bone marrow transplantation and cellular therapies. When combined with SCF and Flt-3 ligand, it promotes the differentiation of HSCs into megakaryocytes, leading to the production of platelets. It has shown promise in emerging cellular therapies for treating sickle cell disease, beta thalassemia, and other blood-related disorders. Moreover, TPO in combination with other cytokines is utilized in the generation of iNK cells, which have the potential to be incorporated into cellular immunotherapies.

Bioinformatics

• Uniprot:
  • NP_000451.1()