Coated antigen:Purified recombinant N antigen.
Conjugate:SARS-CoV-2 N-HRP antigen conjugate (10X concentrate).
1. Diluent 13:Phosphate Buffer Solution (PBS) is a saline buffer composed of a salt conjugate that at 1X concentration consists of:
1.1 Sodium Chloride (NaCl) 1.37×10-7 M
1.2 Potassium chloride (KCl) 2.7×10-9 M
1.3 Dipotassium phosphate (KH2PO4) 2×10-9M
1.4 Disodium phosphate (Na2HPO4) 8×10-9M
2. Positive Control:Inactivated serum containing SARS-CoV-2 antibodies, red color.
3. Negative Control:inactivated non-reactive serum, absent color.
4.2 Tween 80 is a nonionic polyoxyethylene surfactant.
4.3 Fetal bovine serum (FBS) 0.5%:
These components serve the function of facilitating the separation of substances, acting as a humectant, detergent and fungicide, preventing product degradation.
5. Conjugate 1X:Conjugate of purified recombinant protein N antigen labeled to horseradish peroxidase (HRP), this enzyme is widely used since it allows specific activity, does not reduce the conjugation activity and presents functional groups suitable for antigen or antibody binding.
6. Developing solution:Tetramethylbenzidine (TMB) acts as a chromogenic substrate for the detection of HRP activity, this peroxidase catalyzes the conversion of this substrate into a blue colored product.
7. Stop Solution:sulfuric acid 4N (H2SO4), when this strong acid is applied it allows the TMB substrate to stop detection by HRP peroxidase; this product is colored yellow.
These reagents are specified by the ID VET laboratory, where the Indirect Elisa Kit was purchased.