Human Thrombopoietin Quantikine ELISA Kit Best Seller

Name: Human Thrombopoietin Quantikine ELISA Kit
Brand: R&D Systems
SKU: DTP00B
Rating: 5 (2 reviews)

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

(2)

Citations (44)

Product Datasheet / COA / SDS

100% Guarantee

Product Summary
Product Specifications
Sample Values
Precision
Recovery
Linearity
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

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)

View all Thrombopoietin/Tpo ELISA Kits »

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 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

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.

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

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 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. Tissue specific alternate splicing of human 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.

Thrombopoietin receptor (Tpo R), also known as myeloproliferative leukemia protein (c-Mpl), is a type I transmembrane protein that is a member of the hematopoietin/cytokine receptor superfamily. As a consequence of alternative splicing, there are four identified mRNA variants. The functional receptor is encoded by the P isoform of mRNA. The human and mouse receptors share approximately 81% amino acid sequence identity. Tpo R is expressed at low levels in various cell types, including hematopoietic progenitor cells, megakaryocytes and platelets.

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

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human Thrombopoietin Quantikine ELISA Kit

For research use only

Citations
Reviews

⚠ 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.