Canine C-Reactive Protein ELISA Kit From Helica Biosystems, Inc.

C-reactive protein (CRP) is a positive acute phase molecule secreted by the liver in response to internal and external injury. CRP binds to membrane receptors of neutrophils and mononuclear cells, and stimulates the release of proinflammatory cytokines. In dogs, increasing CRP levels were associated with a number of inflammatory, infectious, and post-traumatic conditions, making it a good marker of tissue damage and healing.

The Canine CRP ELISA Kit from Helica Biosystems contains a 96-well plate coated with purified goat anti-dog CRP-IgG, a vial of concentrated conjugate HRP-labeled goat anti-dog CRP-IgG, a vial of high quality CRP standard, one packet of dehydrated wash buffer, a vial of TMB substrate, and a vial of phosphoric acid as stop solution. The assay is highly specific, sensitive and easy to handle; it is stable for one-year when stored at 2-8°C.

To perform the assay, make sure that all the components have reached room temperature before use. In addition to serum and plasma, a number of specimen types (e.g. urine, saliva) can be used in this assay, making it useful in canine medicine, where sometimes it is difficult to collect certain types of samples. If the choice is to use serum or plasma, make sure that diluted samples (1:500) are used. In my analysis I used diluted serum. To accomplish the measurements, 2 µL of sample is enough. If a specimen other than serum or plasma is used, the manufacturer recommends a dilution of 1:2. A standard curve is made using a series of standards made from the standard concentrate provided. 100 µL of diluted sample is added to each well followed by incubation. The plate is washed and the conjugate is added, followed by incubation and washing steps. The chromogen is then added to start the color development, which appears blue in positive reactions. Finally, the reaction is discontinued with a stop solution, which turns the blue color into yellow in positive reactions. The absorbance of the plate is read in a spectrophotometer or plate reader at 450 nm.