Human Thrombopoietin/Tpo Antibody

AB-288-NA Datasheet / COA / SDS

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AB-288-NA

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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

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Cell Proliferation Induced by Thrombopoietin and Neutralization by Human Thrombopoietin Antibody. Recombinant Human Thrombopoietin/Tpo (Catalog # 288-TP) stimulates proliferation in the MO7e human mega-karyocytic 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 concen-trations of Goat Anti-Human Thrombopoietin/Tpo Polyclonal Antibody (Catalog # AB-288-NA). The ND₅₀ is typically 5-10 µg/mL.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 1 mg/mL in sterile PBS.

Reconstitution Buffer Available

Reconstitution Buffer 1 (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.
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

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.
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

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