Complement Classical Pathway ELISA


The Complement Classical Pathway ELISA is an enzyme immunoassay for the qualitative and/or semi-quantitative determination of functional classical complement pathway in human serum, the result shall not be used for clinical diagnosis or patient management. The Eagle Biosciences Human Complement Classical Pathway ELISA Assay kit is for Research Use Only.

Complement Classical Pathway ELISA

Complement Classical Pathway 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: 10 µL
For Research Use Only

Controls Included

Specimen Collection

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 properly handled 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. Do not use sera which are icteric, lipemic and hemolyzed. Heat-inactivated sera cannot be used. Plasma cannot be used.

Assay Background

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 lectin pathway.

Impaired complement activity causes humans to become susceptible to repetitive fulminant or severe infections and may contribute to the development of autoimmune disease. Inappropriate activation of complement contributes to chronic inflammation and tissue injury.

Invitro activation of the complement sequence leads to the consumption of complement components which, in turn, leads to a decrease in their concentration. Thus, the determination of complement proteins or complement activity is used to indicate whether the complement system has been activated by an immunologic and/or pathogenic mechanism. Both functional and immunochemical complement measurements are used to evaluate subjects when a complement-activating disease is suspected or an inherited deficiency is possible. The level of complement activity evaluated by functional assays such as Eagle Biosciences Complement Classical Pathway ELISA Assay Kit takes into account the rate of synthesis, degradation, and consumption of the components and provides a measure of the integrity of the pathways as opposed to immunochemical methods which specifically measure the concentration of various complement components.

Related Products

Complement TCC ELISA Assay Kit
Complement C4d ELISA Kit
Total Complement Functional Screen ELISA

Additional Information

Assay Principle

The Eagle Biosciences Complement Classical Pathway ELISA Assay 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 Complement Classical Pathway ELISA Assay kit, the wells of the microtitre strips are coated with specific activators of the classical pathway. This in combination with sample dilution buffer composition and serum dilution level ensures that only the Classical pathway is activated.

During the incubation of the diluted serum in the wells, complement is activated by the specific coating. The wells are then washed and the amount of C5b-9 complex formed on the plate surface is detected with a specific alkaline phosphatase-labeled antibody to the C5b-9 neoantigen formed during MAC (Membrane Attack Complex) 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).

Package Inserts

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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