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. Tpo promotes the differentiation, proliferation, and maturation of megakaryocytes and their progenitors. Tpo additionally promotes platelet production, aggregation, ECM adhesion, and activation. 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.
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Human Thrombopoietin Quantikine ELISA Kit
R&D Systems | Catalog # DTP00B
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (200 µL), Serum (200 µL), EDTA Plasma (200 µL), Heparin Plasma (200 µL)
Sensitivity
18.5 pg/mL
Assay Range
31.2-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Human Thrombopoietin Quantikine ELISA Kit
The Quantikine Human Tpo Immunoassay is a 4.5 hour solid phase ELISA designed to measure human Tpo in cell culture supernates, serum, and plasma. It contains recombinant human Tpo and antibodies raised against the recombinant factor. Results obtained using natural human Tpo showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural Tpo.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Human
Specificity
Natural and recombinant human Tpo
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Sample Values
Serum/Platelet-poor Plasma - Samples from apparently healthy volunteers were evaluated
for the presence of human Tpo in this assay. No medical histories were available for the donors
used in this study.
ND=Non-detectable
| Sample Type | Mean of Detectable (pg/mL) | % Detectable | Range (pg/mL) |
| Serum (n=38) | 74.2 | 74 | ND-228 |
| Pletelet-poor EDTA plasma (n=34) | 91.8 | 21 | ND-196 |
| Platelet-poor Heparin plasma (n=37) | 82.8 | 19 | ND-168 |
Cell Culture Supernates:
Human peripheral blood cells (1 x 106
cells/mL) were cultured in DMEM supplemented with
5% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and
100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with
10 μg/mL PHA. Aliquots of the culture supernate were removed on days 1 and 5 and assayed
for levels of human Tpo. No detectable levels were observed.
HepG2 human hepatocellular carcinoma cells were cultured in DMEM supplemented with
10% fetal bovine serum. An aliquot of the culture supernate was removed, assayed for human
Tpo, and measured 81.9 pg/mL.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 204 | 579 | 1140 | 203 | 555 | 1070 |
| Standard Deviation | 13.7 | 15.1 | 52.6 | 19.3 | 48.7 | 92.8 |
| CV% | 6.7 | 2.6 | 4.6 | 9.5 | 8.8 | 8.7 |
EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 194 | 512 | 983 | 187 | 517 | 970 |
| Standard Deviation | 17.5 | 16.5 | 26.8 | 16.3 | 39.2 | 63.9 |
| CV% | 9.0 | 3.2 | 2.7 | 8.7 | 7.6 | 6.6 |
Recovery for Human Thrombopoietin Quantikine ELISA Kit
The recovery of Tpo spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 99 | 89-109 |
| Platelet-poor EDTA Plasma (n=4) | 95 | 85-102 |
| Platelet-poor Heparin Plasma (n=4) | 93 | 86-106 |
| Serum (n=4) | 102 | 90-111 |
Linearity
To assess the linearity of the assay, samples spiked with high concentrations of Tpo were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human Thrombopoietin Quantikine ELISA Kit
Human Thrombopoietin/Tpo ELISA Serum/Plasma Standard Curve
Human Thrombopoietin/Tpo ELISA Cell Culture Supernate Standard Curve
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Thrombopoietin/Tpo
Alternate Names
MGDF, MK-CSF, MKCSF, MPLLG, THCYT1, THPO, Tpo
Gene Symbol
THPO
Additional Thrombopoietin/Tpo Products
Product Documents for Human Thrombopoietin Quantikine ELISA Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Thrombopoietin Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
Citations for Human Thrombopoietin Quantikine ELISA Kit
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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