· Circulating immunocomplexes (CIC) are precipitated with polyethylene glycol (PEG) which also precipitates small quantities of non-complex serum constituents.
· After the wash procedure the precipitate is dissolved in a buffer solution and then tested by radial immunodiffusion (RID) for CIC (IgG, IgA, IgM and complement factors C3c and C1q).
· The precipitation ring can be read after 48 hours against a dark background or after staining with color reagent.
· The kit contains 5 x 32 tests in RID CIC plates.
· Control serum, Wash and Dilution buffer, Color reagent, PEG precipitation reagent, Measuring template and Interpretation chart included.
· The evaluation is performed according to enclosed chart.
· Shelf life: 24 months.
· Detection of elevated levels of circulating immunocomplexes as a feature of many disease conditions (e.g. serum sickness, SLE, rheumatoid arthritis, some forms of glomerulonephritis and other inflammations).
· The detection and quantification of CIC in serum has therefore been considered as an adjunct to the diagnosis and management of disease.
Brief assay procedure:
1. Add serum to PEG precipitation reagent and mix well in test tube. Allow test tube to stand in the refrigerator
2. (2 - 4 °C) for 1 - 3 hours
3. Centrifuge at 2 - 4 °C for 20 minutes.
4. Decant supernatant. Invert tube, allow to stand, and absorb residual liquid with absorbent paper towel.
5. Add wash buffer (2 - 4 °C) and vortex to obtain a uniform suspension.
6. Centrifuge at 2 - 4 °C for 20 minutes.
7. Discard supernatant and dry precipitate.
8. Resuspend precipitate in dilution buffer.
9. Incubate in water-bath at 37 °C for 30 minutes.
10. Uncover the plate before sample application and leave it at laboratory temperature for 15 - 30 minutes.
11. Pipette diluted control serum onto application point No. 1 and pipette each of the PEG-treated samples onto application points No. 2 - 16.
12. Seal RID plates and allow to stand on a level surface at room temperature for at least 48 hours without agitating.
13. Evaluate results.