Human Tie-2 Quantikine ELISA Kit

(20 citations)   
  • 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 (100 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
  • Sensitivity
    0.066 ng/mL
  • Assay Range
    0.2 - 10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant human Tie-2
  • 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.
Control Available
QC172, Quantikine Immunoassay Control Set 832 for Human Tie-2 - Please Inquire
Product Summary
The Quantikine Human Tie-2 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Tie-2 in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human Tie-2 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Tie-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 natural human Tie-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, Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean0.762.575.140.822.655.15
Standard Deviation0.040.110.260.070.170.27
CV%5.34.35.18.56.45.2

Recovery

The recovery of Tie-2 spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 91 85-101
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Tie-2 were serially diluted with the Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Tie-2 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Tie-2

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 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-2 cDNA encodes a 1124 amino acid (aa) residue precursor protein with an 18 residue putative signal peptide, a 727 residue extracellular domain and a 354 residue cytoplasmic domain. Tie-2 is a receptor for the angiopoietin (ANG) family: ANG-1, ANG-2, and ANG-3 (mouse)/-4 (human). Ang-2 has been reported to act as an antagonist for Ang-1. Mice engineered to overexpress Ang-2 or to lack Ang-1 or Tie-2 display similar angiogenesis defects.

    • Long Name
      Tyrosine Kinase with Immunoglobulin and Epidermal Growth Factor Homology Domains 2
    • Entrez Gene IDs
      7010 (Human); 21687 (Mouse); 30747 (Zebrafish);
    • Alternate Names
      angiopoietin-1 receptor; CD202b antigen; CD202b; EC 2.7.10; EC 2.7.10.1; hTIE2; p140 TEK; soluble TIE2 variant 1; soluble TIE2 variant 2; TEK tyrosine kinase, endothelial; TEK; Tie2; Tie-2; TIE2CD202b; Tunica interna endothelial cell kinase; Tyrosine-protein kinase receptor TEK; Tyrosine-protein kinase receptor TIE-2; venous malformations, multiple cutaneous and mucosal; VMCM; VMCM1;
    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. 100 µL Standard, Control, or Sample
    6.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
    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.
    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. 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.

    20 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. Tie2 as a novel key factor of microangiopathy in systemic sclerosis
      Authors: F Moritz, J Schniering, JHW Distler, RE Gay, S Gay, O Distler, B Maurer
      Arthritis Res. Ther., 2017;19(1):105.
      Species: Human
      Sample Type: Serum
    2. TIE2-mediated tyrosine phosphorylation of H4 regulates DNA damage response by recruiting ABL1
      Sci Adv, 2016;2(4):e1501290.
      Species: Human
      Sample Type: Cell Lysates
    3. Angiopoietin/Tie2 Dysbalance Is Associated with Acute Kidney Injury after Cardiac Surgery Assisted by Cardiopulmonary Bypass.
      Authors: Jongman R, van Klarenbosch J, Molema G, Zijlstra J, de Vries A, van Meurs M
      PLoS ONE, 2015;10(8):e0136205.
      Species: Human
      Sample Type: Plasma
    4. Clinical outcome, proteome kinetics and angiogenic factors in serum after thermoablation of colorectal liver metastases.
      Authors: Wertenbroek, Marieke, Schepers, Marianne, Kamminga-Rasker, Hannetta, Bottema, Jan T, Muller Kobold, Anneke C, Roelofsen, Han, de Jong, Koert P
      BMC Cancer, 2013;13(1):266.
      Species: Human
      Sample Type: Serum
    5. Circulating angiopoietin-2 and soluble Tie-2 in type 2 diabetes mellitus: a cross-sectional study.
      Authors: Rasul S, Reiter MH, Ilhan A, Lampichler K, Wagner L, Kautzky-Willer A
      Cardiovasc Diabetol, 2011;10(0):55.
      Species: Human
      Sample Type: Plasma
    6. Shedding of the endothelial receptor tyrosine kinase Tie2 correlates with leukemic blast burden and outcome after allogeneic hematopoietic stem cell transplantation for AML.
      Authors: Koenecke C, Kumpers P, Lukasz A, Dammann E, Verhagen W, Gohring G, Buchholz S, Krauter J, Eder M, Schlegelberger B, Ganser A
      Ann. Hematol., 2010;89(5):459-67.
      Species: Human
      Sample Type: Serum
    7. Defective angiogenesis in hypoplastic human fetal lungs correlates with nitric oxide synthase deficiency that occurs despite enhanced angiopoietin-2 and VEGF.
      Authors: Boucherat O, Franco-Montoya ML, Delacourt C, Martinovic J, Masse V, Elie C, Thebaud B, Benachi A, Bourbon JR
      Am. J. Physiol. Lung Cell Mol. Physiol., 2010;298(6):L849-56.
      Species: Human
      Sample Type: Tissue Homogenates
    8. Changes in vascular endothelial growth factor, angiopoietins, and Tie-2 levels with G-CSF stimulation in healthy donors.
      Authors: Serefhanoglu S, Goker H, Buyukasik Y, Turgut M, Sayinalp N, Haznedaroglu IC, Aksu S, Akman U, Ozcebe OI
      Ann. Hematol., 2009;88(7):667-71.
      Species: Human
      Sample Type: Plasma
    9. The angiopoietin/Tie-2 system regulates pericyte survival and recruitment in diabetic retinopathy.
      Authors: Cai J, Kehoe O, Smith GM, Hykin P, Boulton ME
      Invest. Ophthalmol. Vis. Sci., 2008;49(5):2163-71.
      Species: Bovine
      Sample Type: Cell Lysates
    10. Changes in plasma vascular endothelial growth factor, angiopoietins, and their receptors following surgery for breast cancer.
      Authors: Caine GJ, Stonelake PS, Lip GY, Blann AD
      Cancer Lett., 2007;248(1):131-6.
      Species: Human
      Sample Type: Plasma
    11. Significant decrease in angiopoietin-1 and angiopoietin-2 after radical prostatectomy in prostate cancer patients.
      Authors: Caine GJ, Ryan P, Lip GY, Blann AD
      Cancer Lett., 2007;251(2):296-301.
      Species: Human
      Sample Type: Plasma
    12. Marrow angiogenesis-associated factors as prognostic biomarkers in patients with acute myelogenous leukaemia.
      Authors: Lee CY, Tien HF, Hu CY, Chou WC, Lin LI
      Br. J. Cancer, 2007;97(7):877-82.
      Species: Human
      Sample Type: Plasma
    13. Circulating levels of total angiopoietin-2 and the soluble Tie-2 receptor in women during ovarian stimulation and early gestation.
      Authors: Molskness TA, Stouffer RL, Burry KA, Gorrill MJ, Lee DM, Patton PE
      Fertil. Steril., 2006;86(5):1531-3.
      Species: N/A
      Sample Type:
    14. Differential levels of soluble angiopoietin-2 and Tie-2 in patients with haematological malignancies.
      Authors: Quartarone E, Alonci A, Allegra A, Bellomo G, Calabro L, D'Angelo A, Del Fabro V, Grasso A, Cincotta M, Musolino C
      Eur. J. Haematol., 2006;77(6):480-5.
      Species: Human
      Sample Type: Serum
    15. Potential role of soluble angiopoietin-2 and Tie-2 in patients with inflammatory bowel disease.
      Authors: Koutroubakis IE, Xidakis C, Karmiris K, Sfiridaki A, Kandidaki E, Kouroumalis EA
      Eur. J. Clin. Invest., 2006;36(2):127-32.
      Species: Human
      Sample Type: Serum
    16. Differential expression of angiopoietin-1, angiopoietin-2, and Tie receptors in placentas from pregnancies complicated by placenta accreta.
      Authors: Tseng JJ, Hsu SL, Ho ES, Hsieh YT, Wen MC, Chou MM
      Am. J. Obstet. Gynecol., 2006;194(2):564-71.
      Species: Human
      Sample Type: Tissue Homogenates
    17. Angiopoietin 1 (Ang-1), angiopoietin 2 (Ang-2) and Tie-2 (a receptor tyrosine kinase) concentrations in peripheral blood of patients with thyroid cancers.
      Authors: Niedzwiecki S, Stepien T, Kopec K, Kuzdak K, Komorowski J, Krupinski R, Stepien H
      Cytokine, 2006;36(5):291-5.
      Species: Human
      Sample Type: Serum
    18. Phase I clinical and pharmacokinetic study of PTK/ZK, a multiple VEGF receptor inhibitor, in patients with liver metastases from solid tumours.
      Authors: Mross K, Drevs J, Muller M, Medinger M, Marme D, Hennig J, Morgan B, Lebwohl D, Masson E, Ho YY, Gunther C, Laurent D, Unger C
      Eur. J. Cancer, 2005;41(9):1291-9.
      Species: Human
      Sample Type: Serum
    19. Multiple roles for the receptor tyrosine kinase axl in tumor formation.
      Authors: Holland SJ, Powell MJ, Franci C, Chan EW, Friera AM, Atchison RE, McLaughlin J, Swift SE, Pali ES, Yam G, Wong S, Lasaga J, Shen MR, Yu S, Xu W, Hitoshi Y, Bogenberger J, Nör JE, Payan DG, Lorens JB
      Cancer Res., 2005;65(20):9294-303.
      Species: Human
      Sample Type: Cell Lysates
    20. Bi-directional cell contact-dependent regulation of gene expression between endothelial cells and osteoblasts in a three-dimensional spheroidal coculture model.
      Authors: Stahl A, Wenger A, Weber H, Stark GB, Augustin HG, Finkenzeller G
      Biochem. Biophys. Res. Commun., 2004;322(2):684-92.
      Species: Human
      Sample Type: Cell Culture Supernates
    Expand to show all 20 Citations
    ELISA Controls
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    Quantikine Immunoassay Control Set 832 for Human Tie-2

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    Quantikine ELISA Wash Buffer 1

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