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Human Tie-1 Quantikine ELISA Kit

Catalog #: DTE100
1 Citation Datasheet / COA / SDS

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DTE100 has been discontinued.
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Summary Precision Recovery Linearity Product Datasheets Preparation and Storage Background Related Research Areas

Human Tie-1 Quantikine ELISA Kit Summary

Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
Sensitivity
0.021 ng/mL
Assay Range
0.2 - 10 ng/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human Tie-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.

Product Summary

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

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, Cell Lysates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (ng/mL) 1.41 2.55 5.73 1.34 2.42 5.27
Standard Deviation 0.08 0.09 0.26 0.1 0.15 0.23
CV% 5.7 3.5 4.5 7.5 6.2 4.4

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (ng/mL) 1.17 2.23 4.84 1.21 2.2 4.84
Standard Deviation 0.06 0.07 0.16 0.08 0.14 0.17
CV% 5.1 3.1 3.3 6.6 6.4 3.5

Recovery

The recovery of Tie-1 spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 99 90-108

Linearity

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

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: Tie-1

Tie-1/Tie and Tie-2/Tek are receptor tyrosine kinases with unique structural characteristics including two immunoglobulin-like domains flanking three epidermal growth factor (EGF)-like domains, followed by three fibronectin type III-like repeats in the extracellular region, and a split tyrosine kinase domain in the cytoplasmic region. Tie-2 is a receptor for the angiopoietin (ANG) family: ANG-1, ANG-2, and ANG-3 (mouse)/-4 (human). It is involved in vascular stabilization and remodeling. Although less well understood, Tie-1 may also act as an ANG receptor, possibly in complex with Tie-2.

Human Tie-1 cDNA encodes a 1138 amino acid (aa) residue precursor protein with a 24 residue putative signal peptide, a 735 residue extracellular domain and a 354 residue cytoplasmic domain. Ligands which bind and activate Tie-1 have not been identified. Based on gene targeting studies, the in vivo functions of Tie-1 have been shown to be related to endothelial cell differentiation and the maintenance of integrity of the endothelium.

Long Name:
Tyrosine Kinase with Immunoglobulin and Epidermal Growth Factor Homology Domains 1
Entrez Gene IDs:
7075 (Human); 21846 (Mouse)
Alternate Names:
EC 2.7.10; EC 2.7.10.1; JTK14soluble TIE1 variant 3; receptor tyrosine kinase; soluble TIE1 variant 1; soluble TIE1 variant 2; soluble TIE1 variant 5; Tie1; Tie-1; TIEsoluble TIE1 variant 4; tyrosine kinase with immunoglobulin and epidermal growth factor homologydomains 1; tyrosine kinase with immunoglobulin-like and EGF-like domains 1; tyrosine-protein kinase receptor Tie-1

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
  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.
  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.
  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. 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.

Citation for Human Tie-1 Quantikine ELISA Kit

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.

1 Citation: Showing 1 - 1

  1. Intermediate monocytes but not TIE2-expressing monocytes are a sensitive diagnostic indicator for colorectal cancer.
    Authors: Schauer D, Starlinger P, Reiter C, Jahn N, Zajc P, Buchberger E, Bachleitner-Hofmann T, Bergmann M, Stift A, Gruenberger T, Brostjan C
    PLoS ONE, 2012-09-04;7(9):e44450.
    Species: Human
    Sample Types: Plasma

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