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Human Thrombopoietin Quantikine ELISA Kit Best Seller

R&D Systems, part of Bio-Techne | Catalog # DTP00B

R&D Systems, part of Bio-Techne
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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)

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

Reactivity

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.

Sample TypeMean of Detectable (pg/mL)% DetectableRange (pg/mL)
Serum (n=38)74.274ND-228
Pletelet-poor EDTA plasma (n=34)91.821ND-196
Platelet-poor Heparin plasma (n=37)82.819ND-168
ND=Non-detectable

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.

Human Thrombopoietin/Tpo ELISA Linearity

Scientific Data Images for Human Thrombopoietin Quantikine ELISA Kit

Human Thrombopoietin/Tpo ELISA Serum/Plasma Standard Curve

Human Thrombopoietin/Tpo ELISA Serum/Plasma Standard Curve

Human Thrombopoietin/Tpo ELISA Cell Culture Supernate 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

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.

Alternate Names

MGDF, MK-CSF, MKCSF, MPLLG, THCYT1, THPO, Tpo

Entrez Gene IDs

7066 (Human); 21832 (Mouse); 81811 (Rat)

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 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

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⚠ 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.