Human ANG-R-Tie2 (Angiopoietin Receptor Tie2) ELISA Kit (HUES01559)
- Product Type:
- ELISA Kit
- 96 Assays
- ELISA Type:
- TEK, CD202B, TIE-2, TIE2, VMCM, VMCM1
- Tested Sample Types:
- Serum, plasma and other biological fluids
|Detection Range:||0.16-10 ng/mL|
|Sample Volume Required Per Well:||100µL|
|Sample Type:||Serum, plasma and other biological fluids|
|Specificity:||This kit recognizes Human ANG-R-Tie2 in samples. No significant cross-reactivity or interference between Human ANG-R-Tie2 and analogues was observed.|
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human ANG-R-Tie2. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human ANG-R-Tie2 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human ANG-R-Tie2, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human ANG-R-Tie2. The concentration of Human ANG-R-Tie2 in samples can be calculated by comparing the OD of the samples to the standard curve.
|UniProt Protein Function:||TIE2: a receptor tyrosine kinase of the Tie family. Receptor for angiopoietin 1. Expressed almost exclusively in endothelial cells. May constitute the earliest mammalian endothelial cell lineage marker. Appears to be critical for endothelial cell-smooth muscle cell communication in venous morphogenesis. TEK is closely related to the TIE receptor tyrosine kinase.|
|UniProt Protein Details:|
Protein type:Protein kinase, TK; Protein kinase, tyrosine (receptor); EC 2. 7. 10. 1; Kinase, protein; Membrane protein, integral; TK group; Tie family
Chromosomal Location of Human Ortholog: 9p21
Cellular Component: microvillus; focal adhesion; cell surface; basolateral plasma membrane; integral to plasma membrane; extracellular region; actin filament; intercellular junction; lipid raft; perinuclear region of cytoplasm; apical plasma membrane; stress fiber; basal plasma membrane; plasma membrane
Molecular Function:protein binding; growth factor binding; protein-tyrosine kinase activity; receptor activity; transmembrane receptor protein tyrosine kinase activity; ATP binding; protein kinase activity
Biological Process: response to peptide hormone stimulus; peptidyl-tyrosine phosphorylation; response to cAMP; heart development; protein amino acid autophosphorylation; cell-matrix adhesion; positive regulation of cytokine secretion during immune response; signal transduction; cell-cell adhesion; cell-cell signaling; positive regulation of focal adhesion formation; angiogenesis; endochondral ossification; organ regeneration; Tie receptor signaling pathway; regulation of establishment and/or maintenance of cell polarity; positive regulation of phosphoinositide 3-kinase activity; positive regulation of peptidyl-serine phosphorylation; protein oligomerization; positive regulation of phosphoinositide 3-kinase cascade; positive regulation of protein kinase B signaling cascade; negative regulation of angiogenesis; positive regulation of angiogenesis; positive regulation of protein import into nucleus; response to estrogen stimulus; negative regulation of inflammatory response; endothelial cell proliferation; response to hypoxia; positive regulation of endothelial cell proliferation; regulation of vascular permeability; sprouting angiogenesis; positive regulation of protein amino acid phosphorylation; blood coagulation; leukocyte migration; transmembrane receptor protein tyrosine kinase signaling pathway; negative regulation of apoptosis
Disease: Venous Malformations, Multiple Cutaneous And Mucosal
|NCBI Summary:||The TEK receptor tyrosine kinase is expressed almost exclusively in endothelial cells in mice, rats, and humans. This receptor possesses a unique extracellular domain containing 2 immunoglobulin-like loops separated by 3 epidermal growth factor-like repeats that are connected to 3 fibronectin type III-like repeats. The ligand for the receptor is angiopoietin-1. Defects in TEK are associated with inherited venous malformations; the TEK signaling pathway appears to be critical for endothelial cell-smooth muscle cell communication in venous morphogenesis. TEK is closely related to the TIE receptor tyrosine kinase. [provided by RefSeq, Jul 2008]|
|NCBI GenInfo Identifier:||218511853|
|NCBI Gene ID:||7010|
|NCBI Accession:||Q02763. 2|
|UniProt Secondary Accession:||Q02763,Q5TCU2, Q8IV34, A8K6W0, B4DH20, B4DHD3, D3DRK5 E7EWI2,|
|UniProt Related Accession:||Q02763|
|Molecular Weight:||60. 2kD|
|NCBI Full Name:||Angiopoietin-1 receptor|
|NCBI Synonym Full Names:||TEK tyrosine kinase, endothelial|
|NCBI Official Symbol:||TEK|
|NCBI Official Synonym Symbols:||TIE2; VMCM; TIE-2; VMCM1; CD202B|
|NCBI Protein Information:||angiopoietin-1 receptor; hTIE2; p140 TEK; soluble TIE2 variant 1; soluble TIE2 variant 2; endothelial tyrosine kinase; tyrosine-protein kinase receptor TEK; tunica interna endothelial cell kinase; tyrosine-protein kinase receptor TIE-2; tyrosine kinase with Ig and EGF homology domains-2|
|UniProt Protein Name:||Angiopoietin-1 receptor|
|UniProt Synonym Protein Names:||Endothelial tyrosine kinase; Tunica interna endothelial cell kinase; Tyrosine kinase with Ig and EGF homology domains-2; Tyrosine-protein kinase receptor TEK; Tyrosine-protein kinase receptor TIE-2; hTIE2; p140 TEK|
|Protein Family:||Angiopoietin-1 receptor|
|UniProt Gene Name:||TEK|
|UniProt Entry Name:||TIE2_HUMAN|
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human ANG-R-Tie2 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human ANG-R-Tie2 were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||6.00||5.30||3.07||7.41||5.26||3.31|
The recovery of Human ANG-R-Tie2 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||90-105||96|
|Cell culture media (n=5)||95-108||101|
Samples were spiked with high concentrations of Human ANG-R-Tie2 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
|Serum (n=5)||EDTA plasma (n=5)||Cell culture media (n=5)|
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
|Micro ELISA Plate(Dismountable)||8 wells ×12 strips||-20°C, 6 months|
|Reference Standard||2 vials|
|Concentrated Biotinylated Detection Ab (100×)||1 vial, 120 µL|
|Concentrated HRP Conjugate (100×)||1 vial, 120 µL||-20°C(shading light), 6 months|
|Reference Standard & Sample Diluent||1 vial, 20 mL||4°C, 6 months|
|Biotinylated Detection Ab Diluent||1 vial, 14 mL|
|HRP Conjugate Diluent||1 vial, 14 mL|
|Concentrated Wash Buffer (25×)||1 vial, 30 mL|
|Substrate Reagent||1 vial, 10 mL||4°C(shading light)|
|Stop Solution||1 vial, 10 mL||4°C|
|Plate Sealer||5 pieces|
|Product Description||1 copy|
|Certificate of Analysis||1 copy|
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100 µL of standard solutions into the standard wells.
- Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
- Add 100 µL of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37 °C.
- Aspirate the liquid from each well, do not wash. Immediately add 100 µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37 °C.
- Aspirate or decant the solution from the plate and add 350 µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100 µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37 °C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 90 µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37 °C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.