Mouse/Rat Angiopoietin-2 Quantikine ELISA Kit Summary
Cell Culture Supernates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma
|Intra-Assay Precision||Inter-Assay Precision|
The recovery of Angiopoietin-2 spiked to levels throughout the range of the assay in various matrices was evaluated.
|Sample Type||Average % Recovery||Range %|
|Cell Culture Samples (n=4)||114||108-120|
|EDTA Plasma (n=2)||105||101-110|
|Heparin Plasma (n=2)||107||103-111|
Preparation and Storage
Angiopoietins, Ang-1, Ang-2, and Ang-3 (mouse)/Ang-4 (human), are natural agonists or antagonists of the Tie-2 receptor tyrosine kinase and are important modulators of angiogenesis. Two domains characterize the angiopoietin family of proteins: an N-terminal coiled-coil domain that mediates homo-oligomerization, and a C-terminal fibrinogen-like domain that binds Tie-2.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 50 µL of Assay Diluent to each well.
- Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
- Aspirate and wash 4 times.
- Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
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