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Human Thrombospondin-1 Quantikine ELISA Kit

R&D Systems | Catalog # DTSP10

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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (25 µL), Serum (10 µL), Platelet-poor EDTA Plasma (25 µL), Platelet-poor Heparin Plasma (25 µL), Saliva (50 µL), Human Milk (50 µL)

Sensitivity

0.944 ng/mL

Assay Range

7.81-500 ng/mL (Cell Culture Supernates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Saliva, Human Milk)
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Product Summary for Human Thrombospondin-1 Quantikine ELISA Kit

The Quantikine Human Thrombospondin-1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Thrombospondin-1 in cell culture supernates, serum, plasma, saliva, and human milk. It contains NS0-expressed recombinant human Thrombospondin-1 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Thrombospondin-1 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-1.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

Specificity

Natural and recombinant human Thrombospondin-1

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.

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, Human Milk, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Saliva, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (ng/mL) 71.9 142 282 75.8 151 294
Standard Deviation 4.34 9.56 18.0 4.71 8.51 17.6
CV% 6.0 6.7 6.4 6.2 5.6 6.0

Recovery for Human Thrombospondin-1 Quantikine ELISA Kit

The recovery of Thrombospondin-1 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 95 86-101
Platelet-poor EDTA Plasma (n=4) 94 89-99
Platelet-poor Heparin Plasma (n=4) 93 87-104

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Thrombospondin-1 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human Thrombospondin-1 ELISA Linearity

Scientific Data Images for Human Thrombospondin-1 Quantikine ELISA Kit

Human Thrombospondin-1 Quantikine ELISA Kit

Human Thrombospondin-1 Quantikine ELISA Kit

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: Thrombospondin-1

Thrombospondin-1 (TSP-1) is a secreted homotrimeric protein that regulates a wide range of cellular functions including their interactions with other cells and with the extracellular matrix (ECM). TSP-1 counteracts the angiogenic, hypotensive, and anti-thrombotic effects of nitric oxide (NO). It binds and neutralizes VEGF, blocks VEGF R2 signaling on vascular endothelial cells (EC), and destabilizes adhesive contacts between EC. TSP-1 also plays an important role in wound repair and tissue fibrosis by binding latent TGF-beta and inducing release of the active cytokine from the latency associated peptide (LAP). TSP-1 dampens adaptive immune responses by inducing the differentiation of regulatory T cells and inhibiting TCR signaling. In the nervous system, it promotes excitatory synapse formation and supports the integration of neural progenitor cells into the olfactory bulb.

Alternate Names

THBS1, Thrombospondin1, TSP-1

Entrez Gene IDs

7057 (Human); 21825 (Mouse)

Gene Symbol

THBS1

Additional Thrombospondin-1 Products

Product Documents for Human Thrombospondin-1 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 Thrombospondin-1 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.

Citations for Human Thrombospondin-1 Quantikine ELISA Kit

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Protocols

View specific protocols for Human Thrombospondin-1 Quantikine ELISA Kit (DTSP10):

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
  3.   Add 100 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   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.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   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.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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.

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