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C3c ELISA kit [Human]-2 x 96 det. (CAT#: CB-P078-K) Datasheet

Product Type
Kit
Description
The complement system mediates many basic biological functions and participates in the host's resistance to infection, the initiation of inflammatory responses, the processing and clearance of immune complexes, and the regulation of immune responses. There are three pathways for complement activation: the classical pathway is initiated by immune complexes, the lectin pathway is initiated by surface-bound mannan-bound lectins, and the alternative pathway is initiated by all surfaces that are not specifically protected. Each complement pathway produces C3 convertase, a serine protease that cleaves the central complement protein C3 and produces the major cleavage fragment C3b. C3b is an opsonin, a part of one of the main invertases that drive the complement cascade. In the presence of complement, the regulatory molecule C3b can be further degraded into iC3b, C3c, C3dg and C3d. The disadvantage of most complement biomarkers is their short half-life, making it difficult to collect and measure reliable samples. Unlike other C3 fragments, C3c does not bind to other structures such as pathogens, cell surface (receptors) and other plasma proteins. Therefore, C3c is a stable complement biomarker and only appears in the liquid phase without interfering with other C3-based products. The measurement of C3c provides evidence of (uncontrolled) complement activation and can be used as an indicator of the inflammatory state. Complement is a key element of the innate immune system. Inappropriate activation is pathological and leads to, for example, various autoimmune diseases (such as HUS). There are also signs that C3 is related to the cardiovascular system and Parkinson's disease, making C3c a reliable and useful biomarker. C3c ELISA is based on antibodies that are only highly specific to C3c epitopes. It is a simple ELISA that can be used in diseases where complement activation is an element of inflammatory response.
Applications
The human C3c ELISA kit is used for the in vitro quantitative determination of human C3c in serum and plasma.
Principle
Human C3c ELISA kit is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle. The working time is 3 hours. The effective form of the plate with twelve disposable 8-well plates allows free choice of batch size for determination. The samples and standards are incubated in microtiter wells coated with antibodies that recognize human C3c. The biotinylated tracer antibody will bind to the captured human C3c. The streptavidin-peroxidase conjugate will bind the biotinylated tracer antibody. The streptavidin-peroxidase conjugate will react with the substrate tetramethylbenzidine (TMB). The enzyme reaction is stopped by adding oxalic acid. Measure the absorbance at 450nm with a spectrophotometer. Draw a standard curve by plotting the absorbance (linear) relative to the corresponding concentration (logarithm) of the human C3c standard. The concentration of human C3c in a sample run at the same time as the standard can be determined from the standard curve.
Size
2 x 96 det.
Detection Method
Enzyme immunoassay (ELISA) technique
Detection Limit
1.6 to 100 ng/ml
Sensitivity
1.6 ng/ml
Sample
Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Specimen Collection
Blood samples were collected using sterile venipuncture techniques, and serum was obtained using standard procedures. At least 5 ml of whole blood is recommended. Let the blood clot in the serum test tube for 60-65 minutes at room temperature (20-25°C). Centrifuge the blood sample and transfer the cell-free serum to a clean test tube. The serum must be handled correctly to prevent complement activation in vitro. Serum should be frozen in a sealed tube at -70°C or lower for long-term storage or transport on dry ice. The sample should not be frozen and thawed more than once. Do not use jaundice, lipemia and hemolytic serum. You cannot use heat-inactivated serum. Plasma cannot be used.
Sample Volume
100 µl/well
Species Reactivity
Human
Protocol Length
2 hours
Storage
Product should be stored at 4 °C. Under recommended storage conditions, product is stable for at least six months.
Research Use
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 with the use of or derivation of this product.

All products and services are for Research Use Only. Do Not use in humans.

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