Ovalbumin (OVA) ELISA Kit

This assay employs the indirect competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with OVA. Standards or samples are then added to the appropriate microtiter plate wells with a Horseradish Peroxidase (HRP)-conjugated antibody specific for OVA and incubated. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of OVA bound in the initial step. The color development is stopped and the intensity of the color is measured.

Ovalbumin is the main protein found in egg white, making up 60-65% of the total protein. While Ovalbumin displays sequence and three-dimensional homology to the serpin superfamily, it is a noninhibitory serpin. While most serpins control such processes as fibrinolysis and coagulation by inhibiting serine proteases, the function of ovalbumin is unknown, although it is presumed to be a storage protein. The ovalbumin protein of chickens is made up of 385 amino acids, and its relative molecular mass is 45 kDa. It is a glycoprotein with 4 sites of glycosylation. It is secreted from the cell. Unlike the signal sequences of most secreted proteins, Ovalbumin's signal sequence is not cleaved. Additionally it is not located directly on the N-terminus, but rather internally, starting at residue 21 and ending at residue 47.

Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 2 - 8°C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2 - 8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles.

Other biological fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.