Total Complement Functional Screen ELISA
Total Complement Functional Screen ELISA Developed and Manufactured by Svar Life Science
Size: 1×96 wells
Sensitivity: See Package Insert
Dynamic Range: Cut-off
Incubation Time: 2 hours
Sample Type: Serum
Sample Size: 20 µL
For Research Use Only
Blood samples are to be collected using aseptic venipuncture technique and serum obtained using standard procedures. A minimum of 5 mL of whole blood is recommended. Allow blood to clot in serum tubes, for 60-65 minutes at room temperature (20-25° C). Centrifuge blood samples and transfer cell-free serum to a clean tube. Sera must be handled properly to prevent in vitro complement activation. Sera should be frozen at -70° C or lower in tightly sealed tubes for extended storage or for transport on dry ice. Samples should not be frozen and thawed more than once. Avoid using sera which are icteric, lipemic and hemolyzed. Heat-inactivated sera can not be used. Plasma can not be used.
The Complement System Screen Kit 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 Total Complement Functional Screen ELISA Assay Kit the wells of the microtitre strips are coated with specific activators of the classical, or MBL, or the alternative pathway. Subject serum is diluted in diluent containing specific blocker to ensure that only the respective pathway is activated. During the incubation of the diluted subject 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-labeled 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 color intensity and is measured in terms of absorbance (optical density (00)).
Learn more about the Complement System at Eagle Biosciences Biomarker Spotlight page dedicated to the Complement System here: