Human Anti-Coronavirus IgG ELISA
The Human Anti-Coronavirus IgG ELISA is For Research Use Only
Size: 12×8 wells
Sensitivity: 800 pg/ml
Standard Range: 0 ng/ml – 1000 ng/ml
Incubation Time: 3 hours
Sample Type: Respiratory Specimens, serum, plasma, cell culture supernate
Sample Size: 100 µL
Alternative Name: Coronavirus, SARS-CoV
The method employs sandwich ELISA technique. Monoclonal antibodies are pre-coated onto microwells. Samples and standards are pipetted into microwells and Coronavirus present in the sample are bound by the antibodies. HRP labeled anti-Human IgG is added and incubated to form a complex. After washing microwells in order to remove any non-specific binding, the substrate solution (TMB) is added to microwells and color develops proportionally to the amount of Coronavirus in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
Sample Preparation and Storage
Specimens should be clear and non-hemolyzed. Samples should be run at a number of dilutions to ensure accurate quantitation.
Blood is taken by venipuncture. Serum is separated after clotting by centrifugation. Plasma can be used, too. Lipaemic, hemolytic or contaminated samples should not be run. Repeated freezing and thawing should be avoided. If samples are to be used for several assays, initially aliquot samples and keep at – 20°C.
Cell Culture Supernates – Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20℃ or lower temperature. Avoid repeated freeze-thaw cycles. If the use of original supernate sample or low dilution (<5 fold) are necessary due to the expected low concentration of antigen supernates need be adjust to neutral pH condition before assay.
Should you desire to inactivate the virus, use a (5X) Lysis Buffer (optional, not provided). Add 1/5 volume of (5X) Lysis Buffer to sample (i.e. add 50 ul (5X) Lysis Buffer to 200 ul sample). Vortex well.
Note: The sample should be diluted to within the working range of the assay in 1X Assay Diluent. The exact dilution must be determined based on the concentration of specific target in individual samples.
Respiratory Sample – Centrifuge samples for 20 minutes at 10000 x g at 2-8°C. Collect supernatant and carry out the assay immediately.