Human TPO/Thrombopoietin ELISA Kit
CAT.NO. : AEH0197
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Background
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. The 356 amino acid (aa) mouse Tpo precursor is cleaved to yield the 335 aa mature protein. Mature mouse Tpo shares 71% and 81% aa sequence homology with human and rat Tpo, respectively. 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. Tissue specific alternate splicing of mouse Tpo generates multiple isoforms with internal deletions, insertions, and/or C‑terminal substitutions. Tpo promotes the differentiation, proliferation, and maturation of MK and their progenitors. Several other cytokines can promote these functions as well but only in cooperation with Tpo. Notably, IL-3 independently induces MK development, although its effects are restricted to early in the MK lineage. Tpo additionally promotes platelet production, aggregation, ECM adhesion, and activation. It is cleaved by platelet-derived thrombin following Arg191 within the C‑terminal domain and subsequently at other sites upon extended digestion. Full length Tpo and shorter forms circulate in the plasma. The C‑terminal domain is not required for binding to Tpo R or inducing MK growth and differentiation. 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.
Typical data
|
pg/ml |
O.D. |
Average |
Corrected |
|
|
0.00 |
0.0260 |
0.0252 |
0.0256 |
|
|
8.19 |
0.0460 |
0.0467 |
0.0464 |
0.0208 |
|
20.48 |
0.0769 |
0.0723 |
0.0746 |
0.0490 |
|
51.20 |
0.1445 |
0.1427 |
0.1436 |
0.1180 |
|
128.00 |
0.3214 |
0.3069 |
0.3142 |
0.2886 |
|
320.00 |
0.6924 |
0.6953 |
0.6939 |
0.6683 |
|
800.00 |
1.6190 |
1.5460 |
1.5825 |
1.5569 |
|
2000.00 |
3.2320 |
3.2120 |
3.2220 |
3.1964 |
Precision
|
Intra-assay Precision |
Inter-assay Precision |
|||||
|
Sample Number |
S1 |
S2 |
S3 |
S1 |
S2 |
S3 |
|
22 |
22 |
22 |
6 |
6 |
6 |
|
|
Average(pg/ml) |
40.7 |
197.2 |
578.0 |
37.2 |
208.0 |
617.8 |
|
Standard Deviation |
2.9 |
9.9 |
34.8 |
2.4 |
11.7 |
41.3 |
|
Coefficient of Variation(%) |
7.1 |
5.0 |
6.0 |
6.5 |
5.6 |
6.7 |
Inter-assay Precision (Precision between assays) Three samples of known concentration were tested six times on one plate to assess intra-assay precision.
Spike Recovery
The spike recovery was evaluated by spiking 3 levels of human TPO/Thrombopoietin into health human serum sample. The un-spiked serum was used as blank in this experiment.
The recovery ranged from 88% to 128% with an overall mean recovery of 114%.
The recovery ranged from 88% to 128% with an overall mean recovery of 114%.
Sample Values
| Sample Matrix | Sample Evaluated | Range (pg/ml) | Detectable (%) | Mean of Detectable (pg/ml) |
|---|---|---|---|---|
| Serum | 30 | 5.60-1005.88 | 100 | 253.32 |
Serum/Plasma – Thirty samples from apparently healthy volunteers were evaluated for the presence of TPO/Thrombopoietin in this assay. No medical histories were available for the donors.
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