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Human Tie-2 DuoSet ELISA

Catalog #: DY5159
10 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY5159
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Human Tie-2 ELISA Standard Curve
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Product Details
Procedure
Citations (10)
FAQs
Supplemental Products
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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human Tie-2 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
156.0 - 10,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Tie-2. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

N/A Human Tie-2 ELISA Standard Curve View Larger

Human Tie-2 ELISA Standard Curve

Product Datasheets

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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.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Tie-2

Tie-2, also known as Tek, is a transmembrane receptor tyrosine kinase (RTK) that functions as a receptor for Angiopoietin family proteins. It is expressed by embryonic and adult endothelial cells, hematopoietic stem cells, and a circulating population of proangiogenic monocytes. Tie-2 signaling is activated by Angiopoietins-1 and -4, while it can be either activated or inhibited by Angiopoietins-2 and -3. Tie-2 plays an important role in maintaining vascular integrity by mediating endothelial cell-smooth muscle cell communication and inhibiting endothelial cell apoptosis. It is also required for embryonic development of the endocardium. In addition, Tie-2 signaling mediates the quiescence of bone marrow stem cells in response to osteoblast-produced Angiopoietin-1. This quiescence is critical for maintaining an ongoing hematopoietic capability in the bone marrow.

Long Name:
Tyrosine Kinase with Immunoglobulin and Epidermal Growth Factor Homology Domains 2
Entrez Gene IDs:
7010 (Human); 21687 (Mouse); 89804 (Rat); 396729 (Porcine); 403714 (Canine); 102122204 (Cynomolgus Monkey); 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

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human Tie-2 DuoSet ELISA

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.

10 Citations: Showing 1 - 10
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  1. Profiling of selected angiogenesis-related genes in serous ovarian cancer patients
    Authors: P Ku?, R Charkiewic, D Klasa-Mazu, T Milczek, B Mroczko, J Nikli?ski, P Lauda?ski
    Adv Med Sci, 2017-02-21;62(1):116-120.
    Species: Human
    Sample Types: Cell Lysates
  2. Biomarkers of Host Response Predict Primary End-Point Radiological Pneumonia in Tanzanian Children with Clinical Pneumonia: A Prospective Cohort Study.
    Authors: Erdman L, D'Acremont V, Hayford K, Rajwans N, Kilowoko M, Kyungu E, Hongoa P, Alamo L, Streiner D, Genton B, Kain K
    PLoS ONE, 2015-09-14;10(9):e0137592.
    Species: Human
    Sample Types: Plasma
  3. Malaria in Pregnancy Interacts with and Alters the Angiogenic Profiles of the Placenta.
    Authors: Ataide R, Murillo O, Dombrowski J, Souza R, Lima F, Lima G, Hristov A, Valle S, Di Santi S, Epiphanio S, Marinho C
    PLoS Negl Trop Dis, 2015-06-19;9(6):e0003824.
    Species: Human
    Sample Types: Plasma
  4. Host biomarkers distinguish dengue from leptospirosis in Colombia: a case-control study.
    Authors: Conroy A, Gelvez M, Hawkes M, Rajwans N, Liles W, Villar-Centeno L, Kain K
    BMC Infect Dis, 2014-01-20;14(0):35.
    Species: Human
    Sample Types: Serum
  5. Increased endothelial inflammation, sTie-2 and arginase activity in umbilical cords obtained from gestational diabetic mothers.
    Authors: Giri H, Chandel S, Dwarakanath L, Sreekumar S, Dixit M
    PLoS ONE, 2013-12-20;8(12):e84546.
    Species: Human
    Sample Types: Serum
  6. Performance characteristics of combinations of host biomarkers to identify women with occult placental malaria: a case-control study from Malawi.
    Authors: Conroy AL, Liles WC, Molyneux ME
    PLoS ONE, 2011-12-12;6(12):e28540.
    Species: Human
    Sample Types: Plasma
  7. Combinations of Host Biomarkers Predict Mortality among Ugandan Children with Severe Malaria: A Retrospective Case-Control Study.
    Authors: Erdman LK, Dhabangi A, Musoke C, Conroy AL, Hawkes M, Higgins S, Rajwans N, Wolofsky KT, Streiner DL, Liles WC, Cserti-Gazdewich CM, Kain KC
    PLoS ONE, 2011-02-25;6(2):e17440.
    Species: Human
    Sample Types: Plasma
  8. Endothelium-based biomarkers are associated with cerebral malaria in Malawian children: a retrospective case-control study.
    Authors: Conroy AL, Phiri H, Hawkes M, Glover S, Mallewa M, Seydel KB, Taylor TE, Molyneux ME, Kain KC
    PLoS ONE, 2010-12-29;5(12):e15291.
    Species: Human
    Sample Types: Plasma
  9. Secretion of angiogenic growth factors by villous cytotrophoblast and extravillous trophoblast in early human pregnancy.
    Authors: Lash GE, Naruse K, Innes BA, Robson SC, Searle RF, Bulmer JN
    Placenta, 2010-03-24;31(6):545-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Serum levels of angiogenic factors and their prognostic relevance in bladder cancer.
    Authors: Szarvas T, Jager T, Droste F, Becker M, Kovalszky I, Romics I, Ergun S, Rubben H
    Pathol. Oncol. Res., 2008-09-20;15(2):193-201.
    Species: Human
    Sample Types: Serum

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