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Catalog # HK3010

Classical Complement Pathway, Human, Assay

The Human Classical Complement Pathway ELISA kit is to be used for the in vitro quantitative determination of Classical Complement Pathway activity in serum samples.

The ELISA can also be used to determine the effect of inhibitory compounds for the classical pathway.

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Features activity assay:

  • Quantitative measurement of complement activation
  • Working at low serum dilutions resulting in less false negative results
  • High throughput and offers higher reproducibility in comparison to hemolytic assays
  • Assay time less than 2 hours

The complement system is pivotal in the immune response, functioning in both innate and adaptive immunity. It includes various proteins and receptors present in the body, with three activation pathways: classical complement pathway (CP), lectin pathway (LP), and alternative pathway (AP). The CP starts with C1q binding to immune complexes, leading to C1r and C1s activation and eventually forming C4b2b, a C3 convertase. All three pathways converge at C3b formation, initiating the second convertase that cleaves C5 into C5a and C5b, amplifying complement activity.

Assessing complement proteins, or complement activity, is crucial in clinical settings. Traditional assays like hemolysis tests are challenging to standardize. ELISA-based tests face limitations due to high serum dilutions. The HK3010 classical complement pathway activity assay overcomes these issues, making it a valuable tool for identifying complement abnormalities and deficiencies.

Complement inhibitors, used in therapy, demand accurate, standardized, and high-throughput complement activity assessment. This not only screens the system for diseases but also identifies candidates for complement inhibition, essential given the high costs of complement inhibitors. The HK3010 assay measures complement function in serum, utilizing human IgM-coated wells to detect the MAC complex with a C9 neo-epitope antibody. By applying regression analysis and a specific inhibitor to prevent pathway interference, this assay accurately measures complement activity and inhibitory capacity with an initial serum dilution of 40x.

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Functional studies, Immuno assays

The Human Classical Complement Pathway ELISA kit is to be used for the in vitro quantitative determination of Classical Complement Pathway activity in serum samples. The ELISA can also be used to determine the effect of inhibitory compounds for the classical pathway.

The Human Classical Complement Pathway ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent activity assay with a working time of 1 hour and 45 minutes. The efficient format of a plate with twelve disposable 8-well strips allows free choice of batch size for the assay. Samples and standards are incubated in microtiter wells coated with antibodies activating Human Classical Complement Pathway. Peroxidase-conjugated antibody will bind to a neoepitope of the formed C5b-9 complex. Peroxidase-conjugate will react with the substrate, tetramethylbenzidine (TMB). The enzyme reaction is stopped by the addition of oxalic acid. The absorbance at 450 nm is measured with a spectrophotometer. The Human Classical Complement Pathway activity of samples, which are run concurrently with the standards, can be evaluated on OD-data using regression analysis on logistically transformed values. Complement activity can be expressed as percentage of the activity of a positive standard serum

1 x 96 det.
Product type
Working volume
100 µl/well
Cross reactivity
Monkey – Yes, Pig – Yes
Complement Activity Assay
Storage and stability
Product should be stored at 4 °C. Under recommended storage conditions, product is stable for at least six months.
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result from the use or derivation of this product.
Safety Data Sheet Assay Manual
1530 kb
Technical Data

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