Human Tpo Magnetic Luminex® Performance Assay

Name: Human Tpo Magnetic Luminex® Performance Assay
Brand: R&D Systems
SKU: LUHM288
Rating: 5 (1 reviews)

R&D Systems, part of Bio-Techne | Catalog # LUHM288

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Citations (2)

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Luminex High Performance Assays

Luminex High Performance Assays go through rigorous development, validation, and quality control testing to ensure you are getting the best performance out of your multiplex assay. Panels are customizable so you can order a selection of the analytes or the entire panel.

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Key Product Details

Panel:	Luminex Performance Human Cytokine Panel A
Assay Type:	Magnetic bead-based multiplex assay for the Luminex^® MAGPIX^® platform
Format:	96-well microplate and antibody-coated magnetic beads*
Species:	Human
Analytes Detected:	Thrombopoietin/Tpo

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View Full Technical Details for Human Tpo Magnetic Luminex® Performance Assay

Technical Data
Citations / Reviews (1)

Product Features for Human Tpo Magnetic Luminex® Performance Assay

Magnetic Luminex High Performance Assays are flexible bead-based multiplex assays. They allow up to 50 user-defined target analytes to be simultaneously profiled using cell culture supernates, serum, or plasma samples. Some panels are also validated with human milk, saliva, and/or urine samples.

Simultaneously profile Human Thrombopoietin/Tpo with up to 21 analytes of your choice in one sample
Magnetic format allows for easier wash steps
Undergoes similar validation testing as Quantikine® ELISA Kits
Precise and Specific
Requires a small sample volume (<50 µL)
Run your assay in just 3-5 hours
Mass-calibrated standards for consistent results with every new lot of material
Enough reagents for one 96-well plate

Performance Validation for Human Tpo Magnetic Luminex® Performance Assay

Our Luminex High Performance Assays undergo our most extensive validation testing. All Luminex High Performance Assay Panels are validated for use with serum and plasma samples. Some High Performance Assays are additionally validated for cell culture supernatants, milk, saliva, or urine, as indicated on the individual product data sheets. Luminex High Performance Assays are tested for sensitivity (three-quarters the low standard), intra-assay precision, inter-assay precision, and to ensure assay linearity for validated sample types. Recovery values for individual samples in validated sample types are also tested. Validation data for each analyte can be found in the product datasheet.

Kit Components for Human Tpo Magnetic Luminex® Performance Assay

Standard Cocktail(s)
Bead Diluent
Biotin Antibody Diluent
Calibrator Diluent
Wash Buffer
Streptavidin-PE
One flat-bottom 96-well Microplate
Foil Plate Sealers (4)
Mixing Bottle
Standard Value Card**

**A standard curve must be generated each time an assay is run, utilizing values from the Certificate of Analysis provided with each Luminex Assay.

Bead Set Contents

User-mix antibody-coated Magnetic beads in individual vials
User-mixed cocktail of biotinylated detection antibody

Precision

	Intra-Assay Precision			Inter-Assay Precision
Intra-assay Precision (precision within an assay) - Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (precision between assays) - Three samples of known concentration were tested in twenty separate assays to assess precision between assays.
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	517	1343	2693	548	1303	2653
Standard deviation	29	73	143	33	89	219
CV (%)	5.6	5.5	5.3	5.9	6.8	8.3

Recovery

Sample Type	Average % Recovery	Range (%)
Samples containing and/or spiked with human Tpo were evaluated for recovery.
Cell culture supernate	102	90-116
Serum	129	104-156
EDTA plasma	94	77-114
Heparin plasma	107	90-122

Linearity

		Cell culture supernate	Serum	EDTA plasma	Heparin plasma
Samples containing and/or spiked with human Tpo were serially diluted to evaluate assay linearity.
1:2	Average % of Expected	98	112	117	94
1:2	Range (%)	93-106	98-127	88-144	89-103
1:4	Average % of Expected	101	109	128	95
1:4	Range (%)	88-116	92-123	88-182	92-100
1:8	Average % of Expected	105	104	137	96
1:8	Range (%)	87-127	87-116	90-193	88-102

Build Your Assay

Make Your Selections Below to Configure Your Custom Luminex Assay

Assays

Below are kits that are available based on your selections. These selections may be further customized before adding to the cart or emailing the configuration to your purchasing department or local distributor to place your order. Selected analytes may be included in more than one High Performance Assay. All assay options are displayed.

If you run into bead region incompatibilities, send your Luminex code to our team and find out more.

How would you like these analytes mixed?

Premixed

User Mixed

Please select at least one mixing option to see the kits.

Would you like to split based on dilution?

Do not split

Split based on serum

Split based on plasma

Split based on cell culture supernates

Which Luminex Assay is Right For Me?

Assays for the Luminex platform are offered as High Performance Assays or Discovery Assays . The Luminex High Performance Assays are fully validated panels with a focused selection of analytes. They are available as Configurable User Mix and Premixed options or as Fixed Panels. The Luminex Discovery Assays allow for the maximum number of analytes in a multiplex and is supplied as a premixed kit. View a table comparing the features and benefits of these bead-based multiplex assays on rndsystems.com .

More Information About Magnetic Bead-based Multiplex Kits For The Luminex® Platform

Magnetic Luminex High Performance Assays are designed for use with the Luminex MAGPIX CCD Imager. Alternatively, kits can be used with the Luminex 200™ or FLEXMAP 3D, dual laser, flow-based sorting and detection platforms.

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using a Luminex analyzer. One laser is bead-specific and determines which analyte is being detected. A magnet in the analyzer captures and holds the superparaMagnetic microparticles in a monolayer. Two spectrally distinct Light Emitting Diodes (LEDs) illuminate the beads. One LED identifies the analyte that is being detected and the second LED determines the magnitude of the PE-derived signal, which is in direct proportion to the amount of analyte bound. Each well is imaged with a CCD camera. Kits can also be used with Luminex 100/200 or a FLEXMAP 3D dual laser, flow-based systems.

Scientific Data Examples for Human Tpo Magnetic Luminex® Performance Assay

	Quantist software graph of signal intensity versus concentration for Thrombopoietin/Tpo in the Human Cytokine A Luminex Panel

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.

Item	Value
Entrez Gene IDs	7066 (Human); 21832 (Mouse); 81811 (Rat)
Alternate Names	MGDF, MGDFC-mpl ligand, MK-CSF, MKCSF, ML, MPL ligand, MPLLG, MPLLGMGC163194, Megakaryocyte colony-stimulating factor, Megakaryocyte growth and development factor, Myeloproliferative leukemia virus oncogene ligand, THCYT1, THPO, TPOMKCSF, Thrombopoietin, Tpo, megakaryocyte stimulating factor, thrombopoietin nirs variant 1

Preparation and Storage

Shipping	The product is shipped with polar packs. 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.