Feline Leukemia Virus-gp70 antigen ELISA

Description

Intended use of the test kit

The FeLV gp70 enzyme immunoassay (ELISA) kit is designed to detect gp70 antigen in culture supernatants and virus/gp70-concentrates. The kit procedure is based on an indirect solid phase Enzyme Immuno Assay (EIA). The gp70 molecule is bound by monoclonal antibodies attached to the solid phase. Unbound materials are removed by rinsing. A diluted biotinilated anti gp70 serum is then added. After incubation, unbound materials are removed by rinsing. After streptavidin conjugate incubation and rinsing, the substrate is added and the optical density is measured at 45Onm.
To standardize the FeLV-gp70 EIA, a FeLV standard has to be tested. The concentration of FeLV-gp70 in the FeLV standard is 1000 µg/ml.

Principle of the test kit

The test is based on the reaction of FeLV gp70 antibodies with immobilized FeLV antigen. To this end, monoclonal anti gp70 antibodies have been coated to the wells of a 96-well microtiter plate.
The FeLV-gp70 test sample is added to the wells, and is captured by the coated monoclonal antibodies.

• Qualitative
  The sample is added (diluted 1:1) to the wells of the coated plate.
• Quantitative
  The sample also can be titrated using a 3-step dilution, starting with undiluted (> 1:3 > 1:9 > 1:27).

After washing biotinilated anti FeLV-gp70 antibodies are added to the wells and will bind to gp70 molecules which have been caught.
The complex is detected by a streptavidin horseradish peroxidase (HRPO) conjugate. Color reaction in the wells is directly related to the concentration of FeLV-gp70 in the sample.

Protocol

Feline Leukemia Virus-gp70 ELISA