The Virusys Influenza A Nucleoprotein Antigen Capture ELISA kits (IAV-142, IAV-143) are highly sensitive and specific semi-quantitative assays for the detection of Influenza A nucleoprotein in a wide variety of samples. To enhance the analytical utility of the ELISA kits, we have developed the Influenza A Nucleoprotein Antigen Calibration Kit, which permits the generation of a nucleoprotein calibration curve and estimation of nucleoprotein in a sample.

The Influenza A Nucleoprotein Antigen Concentrate supplied in this product was produced using A/Beijing/262/95 (H1N1). The virus was grown in embryonated eggs, detergent-treated to remove the viral envelope and concentrated by centrifugation. Following SDS-PAGE, nucleoprotein concentration was calculated as a percentage of the total protein in the preparation.

Intended Use:

The Virusys Influenza A Nucleoprotein Antigen Calibration Kit is intended for the production of a Nucleoprotein Antigen calibration curve and is designed for use with the Virusys Influenza A Nucleoprotein Antigen Capture ELISA Kits (IAV-142, IAV-143).

Kit Components:

1. Influenza A Nucleoprotein Antigen Concentrate (250 ng/ml) - 0.2 ml . Store at –20 deg. C

2. Influenza A Nucleoprotein Antigen Diluent (1x) - 50 ml. Store at 2-8 deg. C

Procedure:

The Virusys Influenza A Nucleoprotein Antigen Concentrate is supplied at 250 ng/ml and requires dilution prior to use. For the generation of a calibration curve, prepare an initial dilution of 1:50 in the diluent supplied. This will result in a nucleoprotein concentration of 5 ng/ml. Serial 2-fold dilutions should then be prepared from the initial dilution (see chart below). Depending upon the nature of the sample being tested, it may be necessary to prepare dilutions in the diluent/buffer used for the experiment (tissue culture medium, for example).

Test each dilution in duplicate following the product insert for the appropriate Virusys Influenza A Nucleoprotein Antigen Capture ELISA Kit.

To determine the amount of nucleoprotein in a sample, a standard curve is constructed as follows:

1. Calculate the average value for each antigen concentration and then subtract the 0 ng/ml value from each.
2. Plot the resulting data and read the unknown values by extrapolation. Alternatively, enter the data into a calculator or a software program and calculate the sample concentrations by linear regression.

Representative Calibration Curve: