The increased awareness and interest in the complement system has augmented the need for a simple and objective method to assess the function of complement activity.
With the Euro Diagnostica complement assay solution acomplete picture of the complement function is provided.
The Wieslab® COMPL AP330 kit is an enzyme immunoassay for the qualitative determination of functional alternative complement pathway in human serum.
FOR IN VITRO DIAGNOSTIC USE.
The complement system plays an essential role in chronic, autoimmune and infectious disease. There are three pathways of complement activation, namely the classical, the alternative and the MBL pathway.
Impaired complement activity causes humans to become susceptible to repetitive fulminant or severe infections and may contribute to development of autoimmune disease. Inappropriate activation of complement contributes to chronic inflammation and tissue injury.
The Wieslab® Complement assay combines principles of the hemolytic assay for complement activation with the use of labeled antibodies specific for neoantigen produced as a result of complement activation. The amount of neoantigen generated is proportional to the functional activity of complement pathways.
In the Complement system Alternative Pathway kit the wells of the microtitre strips are coated with specific activators of the alternative pathway. Patient serum is diluted in diluent containing specific blocker to ensure that only the alternative pathway is activated. During the incubation of the diluted patient serum in the wells, complement is activated by the specific coating.
The wells are then washed and C5b-9 is detected with a specific alkaline phosphatase labelled antibody to the neoantigen expressed during MAC formation.
After a further washing step, detection of specific antibodies is obtained by incubation with alkaline phosphatase substrate solution. The amount of complement activation correlates with the colour intensity and is measured in terms of absorbance (optical density (OD)).
- One frame with red coloured break-apart wells (12x8) sealed in a foil pack with a desiccation sachet. The wells are coated with LPS.
- 35 mL Diluent AP (Dil AP), labelled red.
- 13 mL conjugate containing alkaline phosphatase-labelled antibodies to C5b-9 (blue colour).
- 13 mL Substrate solution ready to use.
- 30 mL wash solution 30x concentrated.
- 0,2 mL negative control (NC) containing human serum. (to be diluted as for a patient serum sample).
- 0,2 mL positive control (PC) containing freezed dried human serum, see “Reconstitution of positive control”, in the IFU.
All reagents in the kit are ready for use except washing solution and controls. The reagents should be stored at 2-8° C except the positive control. The positive control must be stored at -20°C.
