DRG Anti-CCP Elisa Kit

PRINCIPLE OF THE TEST 
Highly purified cyclic citrullinated vimentin peptides (CCP) is bound to microwells. The determination is based on an indirect enzyme linked immune reaction with the following steps: Specific antibodies in the patient sample bind to the antigen coated on the surface of the reaction wells. After incubation, a washing step removes unbound and unspecifically bound serum or plasma components. Subsequently added enzyme conjugate binds to the immobilized antibody-antigen-complexes. After incubation, a second washing step removes unbound enzyme conjugate. After addition of substrate solution the bound enzyme conjugate hydrolyses the substrate forming a blue coloured product. Addition of an acid stops the reaction generating a yellow endproduct. The intensity of the yellow colour correlates with the concentration of the antibody-antigen-complex and can be measured photometrically at 450 nm.

SPECIMEN COLLECTION, STORAGE AND HANDLING 
Collect whole blood specimens using acceptable medical techniques to avoid hemolysis. 
Allow blood to clot and separate the serum or plasma by centrifugation. 
Test serum should be clear and non-hemolyzed. Contamination by hemolysis or lipemia should be avoided, but does not interfere with this assay. 
Specimens may be refrigerated at 2 °C - 8 °C for up to five days or stored at -20 °C up to six months.
Avoid repetitive freezing and thawing of serum or plasma samples. This may result in variable loss of antibody activity.
Testing of heat-inactivated sera is not recommended.

STORAGE AND STABILITY 
Store test kit at 2 °C - 8 °C in the dark. 
Do not expose reagents to heat, sun, or strong light during storage and usage. 
Store microplate sealed and desiccated in the clip bag provided. 
Shelf life of the unopened test kit is 18 months from day of production. Unopened reagents are stable until expiration of the kit. See labels for individual batch.
Diluted Wash Buffer and Sample Buffer are stable for at least 30 days when stored at 2 °C - 8 °C. We recommend consumption on the same day.

CALCULATION OF RESULTS
For quantitative results plot the optical density of each calibrator versus the calibrator concentration to create a calibration curve. The concentration of patient samples may then be estimated from the calibration curve by interpolation. Using data reduction software a 4-Parameter-Fit with lin-log coordinates for optical density and concentration is the data reduction method of choice.