|Size||96wells/kit, with removable strips.|
|Sample Type||cell culture supernates, cell lysates, serum and plasma (heparin, EDTA).|
|Product Name||Human TIE2 PicoKine™ ELISA Kit|
|Storage & Handling||Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)|
|Size||96wells/kit, with removable strips.|
|Description||Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human TIE2. 96wells/kit, with removable strips.|
|Cite This Product||Human TIE2 PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0519)|
|Sample Type||cell culture supernates, cell lysates, serum and plasma (heparin, EDTA)..
Anticoagulant(s): heparin or EDTA
*The recommended anticoagulants are proven to not block the antibody binding sites on the target antigen. Please do not collect blood sample with other anticoagulants thata are not specified above or contact us to check for feasibility.
|Immunogen||Expression system for standard: NSO; Immunogen sequence: A23-K745|
|Cross Reactivity||There is cross-reactivity with human TIE1< 0.1%.|
|Antibody Clonalities||Capture Antibody | Detection Antibody:
monoclonal antibody from mouse|polyclonal antibody from goat
|EK0519-CAP||96-well plate precoated with anti-Human TEK antibody||1|
|EK0519-ST||lyophilized recombinant Human TEK standard||10ng/tube|
|EK0519-DA||biotinylated anti-Human TEK antibody||130ul|
|AR1106-1||sample diluent buffer||30ml|
|AR1106-2||antibody diluent buffer||12ml|
|AR1106-3||ABC diluent buffer||12ml|
|AR1104||TMB color developing agent||10ml|
|AR1105||TMB stop solution||10ml|
Materials Required But Not Provided
- Microplate reader in standard size.
- Automated plate washer.
- Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
- Clean tubes and Eppendorf tubes.
- Washing buffer (neutral PBS or TBS).
- Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g NaCl; 450μl of purified acetic acid or 700μl of concentrated hydrochloric acid to 1000ml H2
- Preparation of 0.01 M PBS: Add 8.5g sodium chloride, 1.4g Na O and adjust pH to 7.2-7.
Typical Data Obtained from Human TIE2 PicoKine™ ELISA Kit
(TMB reaction incubate at 37°C for 25-30min)
Intra/Inter Assay Precision
|Intra-Assay Precision||Inter-Assay Precision|
Three samples with differing target protein concentrations were assayed using four different lots to measure the CV% lot to lot variance.
To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots.
|Lots||Lot1 (pg/ml)||Lot2 (pg/ml)||Lot3 (pg/ml)||Lot4 (pg/ml)||Mean (pg/ml)||Standard Deviation||CV (%)|
*The typical data is obtained from Boster's internal QC result and for reference only. It may differ from the lab test results of the end users. It is more important that the user's lab test results reflect the same linearity demonstrated in the typical data than achieving exactly the same O.D. values.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Angiopoietin-1 receptor|
|Tissue Specificity||Detected in umbilical vein endothelial cells. Proteolytic processing gives rise to a soluble extracellular domain that is detected in blood plasma (at protein level). Predominantly expressed in endothelial cells and their progenitors, the angioblasts. Has been directly found in placenta and lung, with a lower level in umbilical vein endothelial cells, brain and kidney. .|
|Alternative Names||Angiopoietin-1 receptor;184.108.40.206;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;CD202b;TEK;TIE2, VMCM, VMCM1;|
|Subcellular Localization||Cell membrane; Single-pass type I membrane protein. Cell junction. Cell junction, focal adhesion. Cytoplasm, cytoskeleton. Secreted. Recruited to cell-cell contacts in quiescent endothelial cells. Colocalizes with the actin cytoskeleton and at actin stress fibers during cell spreading. Recruited to the lower surface of migrating cells, especially the rear end of the cell. Proteolytic processing gives rise to a soluble extracellular domain that is secreted.|
|Molecular Weight||125830 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Tyrosine-protein kinase that acts as cell-surface receptor for ANGPT1, ANGPT2 and ANGPT4 and regulates angiogenesis, endothelial cell survival, proliferation, migration, adhesion and cell spreading, reorganization of the actin cytoskeleton, but also maintenance of vascular quiescence. Has anti-inflammatory effects by preventing the leakage of proinflammatory plasma proteins and leukocytes from blood vessels. Required for normal angiogenesis and heart development during embryogenesis. Required for post- natal hematopoiesis. After birth, activates or inhibits angiogenesis, depending on the context. Inhibits angiogenesis and promotes vascular stability in quiescent vessels, where endothelial cells have tight contacts. In quiescent vessels, ANGPT1 oligomers recruit TEK to cell-cell contacts, forming complexes with TEK molecules from adjoining cells, and this leads to preferential activation of phosphatidylinositol 3-kinase and the AKT1 signaling cascades. In migrating endothelial cells that lack cell-cell adhesions, ANGT1 recruits TEK to contacts with the extracellular matrix, leading to the formation of focal adhesion complexes, activation of PTK2/FAK and of the downstream kinases MAPK1/ERK2 and MAPK3/ERK1, and ultimately to the stimulation of sprouting angiogenesis. ANGPT1 signaling triggers receptor dimerization and autophosphorylation at specific tyrosine residues that then serve as binding sites for scaffold proteins and effectors. Signaling is modulated by ANGPT2 that has lower affinity for TEK, can promote TEK autophosphorylation in the absence of ANGPT1, but inhibits ANGPT1-mediated signaling by competing for the same binding site. Signaling is also modulated by formation of heterodimers with TIE1, and by proteolytic processing that gives rise to a soluble TEK extracellular domain. The soluble extracellular domain modulates signaling by functioning as decoy receptor for angiopoietins. TEK phosphorylates DOK2, GRB7, GRB14, PIK3R1; SHC1 and TIE1. .|
|Research Areas||Angiogenesis, Cancer, Cardiovascular, Developmental Biology, Oncoproteins, Oncoproteins/Suppressors, Organogenesis, Protein Phosphorylation, Receptor Tyrosine Kinases, Signal Transduction, Stem Cells, Tyrosine Kinases, Vasculature
*You can search these to find other products in these research areas.
|Background||Tyrosine kinase with Ig and EGF homology domain 2(Tie-2), also called TEK tyrosine kinase, endothelial(TEK). Tie-2 and tie-1 are expressed in early embryonic vascular system and in maternal decidual vascular endothelial cells, where the vasculature undergoes an active angiogenesis. Tie-2, but not tie-1, expression was also detected in extraembryonic mesoderm of the amnion. Angiogenesis is coordinated with follicular cell growth in goitrogenesis. The angiopoietins, Ang-1 and Ang-2, are angiogenic growth factors acting through Tie-2. Tie-2 and Ang-1 are expressed in thyroid epithelial and endothelial cells, and Tie-2 is regulated by TSH and cAMP in follicular cells. And Tie-2 expression is increased in goiter in both humans and rats, consistent with a role in goitrogenesis. Tie2/Ang-1 signaling pathway plays a critical role in the maintenance of HSCs in a quiescent state in the BM niche. And the Tie-2 signaling pathway is also critical for endothelial cell-smooth muscle cell communication in venous morphogenesis.|
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• Add tissue homogenates into the wells and then add ABC and TMB without adding any biotinylated detection antibody to see if any signal will be observed.
• If no signal is produced, then you can work on the tissue sample by using the kit.
The 1XTBS can be used if the pH value falls in the range of 7.2-7.6.
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