Human Thrombopoietin/Tpo Antibody

Catalog # Availability Size / Price Qty
AB-288-NA
Cell Proliferation Induced by Thrombopoietin and Neutralization by Human Thrombopoietin Antibody.
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Human Thrombopoietin/Tpo Antibody Summary

Species Reactivity
Human
Specificity
Detects human Thrombopoietin/Tpo in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 5% cross‑reactivity with recombinant mouse Tpo is observed.
Source
Polyclonal Goat IgG
Purification
Protein A or G purified
Immunogen
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human Thrombopoietin/Tpo
Ser22-Gly353
Accession # P40225
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Endotoxin Level
<0.01 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
Recombinant Human Thrombopoietin/Tpo (Catalog # 288-TP)
Neutralization
Measured by its ability to neutralize Thrombopoietin/Tpo-induced proliferation in the MO7e human megakaryocytic leukemic cell line. Avanzi, G. et al. (1988) Br. J. Haematol. 69:359. The Neutralization Dose (ND50) is typically 5‑10 µg/mL in the presence of 10 ng/mL Recombinant Human Thrombopoietin/Tpo.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Example

Neutralization Cell Proliferation Induced by Thrombopoietin and Neutralization by Human Thrombopoietin Antibody. View Larger

Cell Proliferation Induced by Thrombopoietin and Neutralization by Human Thrombopoietin Antibody. Recombinant Human Thrombopoietin/Tpo (Catalog # 288-TP) stimulates proliferation in the MO7e human mega­karyocytic leukemic cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human Thrombopoietin/Tpo (10 ng/mL) is neutralized (green line) by increasing concen­trations of Goat Anti‑Human Thrombopoietin/Tpo Polyclonal Antibody (Catalog # AB-288-NA). The ND50 is typically 5-10 µg/mL.

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 1 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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.
  • 6 months, -20 to -70 °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). 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; MK-CSF; ML; MPL ligand; MPLLGMGC163194; Myeloproliferative leukemia virus oncogene ligand; THPO; thrombopoietin nirs variant 1; Thrombopoietin; Tpo; TPOMKCSF

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Isotype Controls

Reconstitution Buffers

Secondary Antibodies

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