TIE2 ELISA Kit (Human) (OKBB00257)

Catalog No: OKBB00257

Size:96 Tests

Human TEK ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Human
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's human TIE2 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for TIE2 has been precoated onto 96-well plates. Standards(NS0, A23-K745) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for TIE2 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the human TIE2 amount of sample captured in plate.
• ELISA Kit Range: 156 pg/ml - 10,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (ng/mL)	0.87	2.75	6.3	1.01	2.88	7.2
  St. Dev.	0.048	0.132	0.334	0.088	0.19	0.422
  %CV	5.51	4.8	5.30	8.71	6.59	5.86

• ELISA Kit Component:
  • Lyophilized recombinant human TEK standard: 10 ng/tube x 2
  • Anti-human TEK Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human TEK antibody: 130 uL, dilution 1:100
  • Antibody diluent buffer: 12 mL
  • Avidin-Biotin-Peroxidase Complex (ABC): 130 uL, dilution 1:100
  • ABC diluent buffer: 12 mL
  • TMB color developing agent: 10 mL
  • TMB stop solution: 10 mL
• Additional Information: Range: 156pg/ml-10, 000pg/ml
  Principle: Aviva’s human TIE2 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human TIE2 specific-specific polyclonal antibodies were precoated onto 96-well plates. The human specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human TIE2 amount of sample captured in plate.
  Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot
  experiment using standards and a small number of samples is recommended.
  2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
  3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
  4. Duplicate well assay is recommended for both standard and sample testing.
  5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
  6. Don’t reuse tips and tubes to avoid cross contamination.
  7. To avoid to use the reagents from different batches together.
  8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the
  marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before
  using.
  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.1 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.2 Tie2/Ang-1 signaling pathway plays a critical role in the maintenance of HSCs in a quiescent state in the BM niche.3 And the Tie-2 signaling pathway is also critical for endothelial cell-smooth muscle cell communication in venous morphogenesis.4
• Reconstitution and Storage: Store at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
• Sample Type: cell culture supernates, serum and plasma (heparin, EDTA)
• Sensitivity: <5 pg/ml
• Predicted Homology Based on Immunogen Sequence: Cross-reactivates with human TIE1< 0.1%.
• Reference: 1. Sato, T. N.; Qin, Y.; Kozak, C. A.; Audus, K. L. : tie-1 and tie-2 define another class of putative receptor tyrosine
  kinase genes expressed in early embryonic vascular system. Proc. Nat. Acad. Sci. 90: 9355-9358, 1993.
  2. Ramsden, J. D.; Cocks, H. C.; Shams, M.; Nijjar, S.; Watkinson, J. C.; Sheppard, M. C.; Ahmed, A.; Eggo, M. C. :
  Tie-2 is expressed on thyroid follicular cells, is increased in goiter, and is regulated by thyrotropin through cyclic
  adenosine 3-prime,5-prime-monophosphate. J. Clin. Endocr. Metab. 86: 2709-2716, 2001.
  3. Arai, F.; Hirao, A.; Ohmura, M.; Sato, H.; Matsuoka, S.; Takubo, K.; Ito, K.; Koh, G. Y.; Suda, T. : Tie2/Angiopoietin-1
  signaling regulates hematopoietic stem cell quiescence in the bone marrow niche. Cell 118: 149-161, 2004.
  4. Vikkula, M.; Boon, L. M.; Carraway, K. L., III; Calvert, J. T.; Diamonti, A. J.; Goumnerov, B.; Pasyk, K. A.; Marchuk, D.
  A.; Warman, M. L.; Cantley, L. C.; Mulliken, J. B.; Olsen, B. R. : Vascular dysmorphogenesis caused by an activating
  mutation in the receptor tyrosine kinase TIE2. Cell 87: 1181-1190, 1996.
• Specificity: Natural and recombinant human TIE2
• Immunogen: Expression system for standard: NS0; Immunogen sequence: A23-K745
• Description: For quantitative detection of human TIE2 in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Target Info

• Gene Symbol: TEK
• Gene Full Name: TEK tyrosine kinase, endothelial
• Alias Symbols: Angiopoietin-1 receptor, CD202b, CD202b antigen, EC 2.7.10.1, hTIE2, p140 TEK, soluble TIE2 variant 1, soluble TIE2 variant 2, Tek, TEK tyrosine kinase, endothelial, TIE 2, TIE2_HUMAN, Tunica interna endothelial cell kinase, Tyrosine-protein kinase receptor TEK, Tyrosine-protein kinase receptor TIE 2, Tyrosine-protein kinase receptor TIE-2, venous malformations, multiple cutaneous and mucosal, VMCM, VMCM1
• NCBI Gene Id: 7010
• Protein Name: Angiopoietin-1 receptor
• Description of Target: 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.i 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.ii Tie2/Ang-1 signaling pathway plays a critical role in the maintenance of HSCs in a quiescent state in the BM niche.iii And the Tie-2 signaling pathway is also critical for endothelial cell-smooth muscle cell communication in venous morphogenesis.iv
• Uniprot ID: Q02763
• Protein Accession #: NP_000450.2
• Nucleotide Accession #: NM_000459.4