Human Endoglin/CD105 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human Endoglin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Endoglin in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human Endoglin and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Endoglin showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring Endoglin.

Preparation and Storage

Background: Endoglin/CD105

Endoglin (CD105) is a transmembrane type III receptor for TGF-beta superfamily ligands and plays an important role in smooth muscle differentiation, angiogenesis, and neovascularization. It is highly expressed on proliferating vascular endothelial cells, chondrocytes, and syncytiotrophoblasts of term placenta. Human Endoglin haploinsufficiency can a cause the vascular disorder, hereditary hemorrhagic telangiectasia type I. Elevated levels of anti-angiogenic soluble Endoglin contribute to pathogenicity in preeclampsia. Endoglin associates with the receptors TGF-beta RII, Activin RIIA or RIIB, BMPR-IA/ALK-3, or BMPR-IB/ALK6 and enhance binding of Activin A, BMP-2, -7, -9, TGF-beta 1, or TGF-beta 3. Endoglin can either enhance or inhibit signaling through these receptor complexes.

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1.     Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2.     Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3.     Add 50 μL of Assay Diluent to each well.

4.     Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5.     Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6.     Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7.     Aspirate and wash 5 times.

8.     Add 100 μL Substrate Solution to each well.

9.     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.