ANGIOTENSIN CONVERTING ENZYME 2 ELISA KIT - (ACE2 No: ACE2-ELISA)

• Direct sandwich ELISA format providing convenience and high sensitivity.
• Monoclonal antibody coated plate and polyiclonal detection antibody
• Strip plate format allowing separate use and storage of ELISA plate sections.
• Rapid protocol:  Only one incubation.  Total protocol time approximately 1.5 hrs.
• Colormetric detection at 450 nm.
• ACE2 is the primary receptor of COVID-19.

Protocol

 

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1. 8 x 12 well removable section strip plate coated with anti-ACE2 monoclonal antibody
2. Anti-ACE2 polyclonal detection antibody conjugated with biotin
3. Steptavidin-HRP
4. Protein Standard
5. Standard diluent
6. Concentrated wash solution
7. Chromogen solutions A and B
8. Stop solution
9. Plate sealers

Background

Angiotensin-converting enzyme 2 (ACE2) is a single pass transmebraneprotein expressed in endothelial and other cells. There is an enzymatically active domain exposed on the surface of cells in lungs and other tissues. The extracellular domain of ACE2 is cleaved from the transmembrane domain by a sheddase, and the resulting soluble protein is released into the blood stream and ultimately excreted into urine.

ACE2 serves as the main entry point into cells for some coronaviruses, including SARS-CoV-2. The binding of the spike S1 protein of SARS-CoV-2 to the enzymatic domain of ACE2 on the surface of cells results in endocytosis and translocation of both the virus and the enzyme into endosomes located within cells. 

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Methods Overview

 

The included ELISA micro-strip plate is provided pre-coated with an a monoclonal capture antibody selective to the antigen.  Samples are pre-mixed with a biotin-linked detection antibody and streptavidin-HRP, which are then added to the plate.  The plate is washed after a 60 min incubation period.  Color reagents are added, resulting in a reaction with HRP that yields a blue colored solution.  A stop solution is applied to terminate the reaction, and the solution is measured at 450nm.  

   Sandwich ELISA Methods                                                                                                                                        

ELISA Assay Steps

1. Mix sample, biotinylated-detection antibody and streptavidin-HRP.
2. Apply sample and antibody-HRP complex to ELISA plate for 60min at 37^oC.  Wash 5 times.
3. Apply chromogens, incubate, then stop reaction.