Human Angiopoietin-2 ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Angiopoietin-2 is a protein that in humans is encoded by the ANGPT2 gene. Angiopoietins are members of the vascular endothelial growth factor family and the only known growth factors largely specific for vascular endothelium. ANGPT2 is a naturally occurring antagonist of angiopoietin-1 that competes for binding to the TIE2 receptor and blocks ANGPT1-induced TIE2 autophosphorylation during vasculogenesis. Its expression occurs at sites of vascular remodelling: dorsal aorta and major aortic branches, ovary, placenta and uterus.

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 327 1634 5900 341 1769 5438 Standard deviation 17.33 114.38 354 23.52 127.36 342.59 CV (%) 5.3% 7% 6% 6.9% 7.2% 6.3% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 327 1634 5900 341 1769 5438 Standard deviation 17.33 114.38 354 23.52 127.36 342.59 CV (%) 5.3% 7% 6% 6.9% 7.2% 6.3% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 327 1634 5900 341 1769 5438 Mean (pg/ml) 327 1634 5900 341 1769 5438 Standard deviation 17.33 114.38 354 23.52 127.36 342.59 Standard deviation 17.33 114.38 354 23.52 127.36 342.59 CV (%) 5.3% 7% 6% 6.9% 7.2% 6.3% CV (%) 5.3% 7% 6% 6.9% 7.2% 6.3%

Binds to TEK/TIE2, competing for the ANGPT1 binding site, and modulating ANGPT1 signaling. Can induce tyrosine phosphorylation of TEK/TIE2 in the absence of ANGPT1. In the absence of angiogenic inducers, such as VEGF, ANGPT2-mediated loosening of cell-matrix contacts may induce endothelial cell apoptosis with consequent vascular regression. In concert with VEGF, it may facilitate endothelial cell migration and proliferation, thus serving as a permissive angiogenic signal.

Catalog number Description Quantity Storage of opened/reconstituted material EZ0657-CA Rabbit anti- human ANGPT2 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0657-DA Biotinylated goat anti- human ANGPT2 polyclonal antibody (Detection Antibody) 500 μl, 7 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0657-ST Lyophilized recombinant human ANGPT2 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ0657-CA Rabbit anti- human ANGPT2 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0657-DA Biotinylated goat anti- human ANGPT2 polyclonal antibody (Detection Antibody) 500 μl, 7 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0657-ST Lyophilized recombinant human ANGPT2 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ0657-CA Rabbit anti- human ANGPT2 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0657-CA Rabbit anti- human ANGPT2 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0657-DA Biotinylated goat anti- human ANGPT2 polyclonal antibody (Detection Antibody) 500 μl, 7 μg/mL (recommended dilution 1:100) EZ0657-DA Biotinylated goat anti- human ANGPT2 polyclonal antibody (Detection Antibody) 500 μl, 7 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0657-ST Lyophilized recombinant human ANGPT2 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EK0657-ST Lyophilized recombinant human ANGPT2 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.095 0.178 0.21 0.376 0.519 0.97 1.588 2.01 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.095 0.178 0.21 0.376 0.519 0.97 1.588 2.01 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.095 0.178 0.21 0.376 0.519 0.97 1.588 2.01 O.D. 0.095 0.178 0.21 0.376 0.519 0.97 1.588 2.01

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)