The Virusys Adenovirus Hexon Antigen Capture ELISA kits (AK290-2 and AK290-5) are highly sensitive and specific semi-quantitative assays for the detection of adenovirus hexon antigen in a wide variety of sample matrices. To enhance the analytical utility of the ELISA kits, Virusys has developed the Adenovirus Hexon Antigen Calibration Kit, which permits the generation of a hexon antigen calibration curve and estimation of hexon antigen concentration in a sample.
The Virusys Adenovirus Hexon Antigen Calibration Kit is intended for the production of an hexon antigen calibration curve and is designed for use with the Virusys Adenovirus Hexon Antigen Capture ELISA Kits (AK290-2 and AK290-5).
Adenovirus Hexon Antigen (Ad5) Concentrate (125 ng/ml) - 0.4ml Store at 2-8° C
Adenovirus Hexon Antigen Diluent (1x) - 1 x 50 ml
Store at 2-8° C
The Virusys Adenovirus Hexon Antigen Concentrate is supplied at a concentration of 125 ng/ml and requires dilution prior to use. For the generation of a calibration curve, prepare an initial dilution of 1:25 in the diluent supplied. This will result in an Ad5 hexon 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 calibrator dilutions in the diluent/buffer used for the experiment (tissue culture medium, for example).
|Hexon (ng/ml)||Hexon Concentrate||Diluent|
|5||40 ul @ 125 ng/ml||960 ul|
|2.5||500 ul @ 5 ng/ml||500 ul|
|1.25||500 ul @ 2.5 ng/ml||500 ul|
|0.625||500 ul @ 1.25 ng/ml||500 ul|
|0.313||500 ul @ 0.625 ng/ml||500 ul|
|0.156||500 ul @ 0.313 ng/ml||500 ul|
|0.078||500 ul @ 0.156 ng/ml||500 ul|
Test each dilution in duplicate following the product insert for the appropriate Virusys Adenonvirus Hexon Antigen Capture ELISA Kit.
To determine the amount of hexon in a sample, a standard curve is constructed as follows:
Calculate the average value for each antigen concentration and then subtract the 0 ng/ml value from each.
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: