Human Thrombospondin-2 Quantikine ELISA Kit

  (2 citations)     
Datasheet / CoA / SDS
Product Details
Assay Procedure
Citations (2)
FAQs
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (13 uL), EDTA Plasma (13 uL), Heparin Plasma (13 uL), Human Milk (50 uL)
  • Sensitivity
    0.068 ng/mL
  • Assay Range
    0.3 - 20 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Human Milk)
  • Specificity
    Natural and recombinant human Thrombospondin-2
  • Cross-reactivity
    Cross-reactivity observed with 1 or more available related molecules.Cross-species reactivity not tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC171 , Quantikine Immunoassay Control Set 841 Hum Thrombospondin-2 - Please Inquire
Product Summary
The Quantikine Human Thrombospondin-2 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Thrombospondin-2 in cell culture supernates, serum, plasma, and human milk. It contains NS0-expressed recombinant human Thrombospondin-2 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Thrombospondin-2 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human Thrombospondin-2.

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 4.08 7.73 10.9 4.11 7.71 11.4
Standard Deviation 0.189 0.229 0.281 0.295 0.511 0.726
CV% 4.6 3 2.6 7.2 6.6 6.4

Serum, EDTA Plasma, Heparin Plasma, Human Milk
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 3.97 7.96 11.4 4.12 7.68 11.4
Standard Deviation 0.198 0.205 0.284 0.259 0.403 0.527
CV% 5 2.6 2.5 6.3 5.2 4.6

Recovery

The recovery of Thrombospondin-2 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 101 91-108
EDTA Plasma (n=4) 93 86-105
Heparin Plasma (n=4) 92 86-103
Serum (n=4) 93 86-105
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Thrombospondin-2 were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
 Thrombospondin-2 [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Thrombospondin-2
Thrombospondins (TSP) are secreted multidomain glycoproteins with many putative functions including modulating cell adhesion, proliferation, migration, and angiogenesis. At least five TSPs exist: TSP-1, -2, -3, -4, and cartilage oligomeric glycoprotein (COMP)/TSP-5. The family can be divided into two subgroups. The first, TSP-1 and -2, are homotrimeric molecules, each monomer containing an N-terminal/heparin-binding domain, a coiled-coil oligomerization domain, a von Willebrand factor-type/procollagen homology domain, three type 1 thrombospondin repeats (TSR), three EGF-like repeats, seven TSP type 3 repeats, and a C-terminal globular domain. TSP-1 and -2 are thought to mediate interactions between cells and the extracellular matrix, and as such are matricellular proteins. In contrast, TSP-3, -4, and COMP/TSP-5 are homopentamers that lack the N-terminal structure of TSP-1 and -2, but have a similar arrangement of EGF and TSP type 3 repeats, and the C-terminal domain characteristic of the TSP family.
  • Entrez Gene IDs:
    7058 (Human)
  • Alternate Names:
    THBS2; thrombospondin 2; Thrombospondin2; Thrombospondin-2; TSP-2; TSP2thrombospondin-2
Related Research Areas
Assay Procedure
Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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Species
Sample Type
  1. Circulating thrombospondin-2 reflects disease severity and predicts outcome of heart failure with reduced ejection fraction.
    Authors: Hanatani S, Izumiya Y, Takashio S, Kimura Y, Araki S, Rokutanda T, Tsujita K, Yamamoto E, Tanaka T, Yamamuro M, Kojima S, Tayama S, Kaikita K, Hokimoto S, Ogawa H
    Circ J, 2014;78(4):903-10.
    Species: Human
    Sample Type: Serum
  2. Tumor-derived pancreatic stellate cells promote pancreatic cancer cell invasion through release of thrombospondin-2.
    Authors: Farrow B, Berger DH, Rowley D
    J. Surg. Res., 2009;156(1):155-60.
    Species: Human
    Sample Type: Cell Culture Supernates

FAQs

  1. What is the normal appearance of Assay Diluent RD1-75?

    • Assay Diluent RD1-75 contains high concentrations of additives and typically has a very cloudy, brown appearance. It is recommended to mix this diluent well before and during use.

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ELISA Controls
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Quantikine Immunoassay Control Set 841 Hum Thrombospondin-2

Ctrl QC171
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Supplemental ELISA Products
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Quantikine ELISA Wash Buffer 1

ELISA WA126 6  
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