Human C3 ELISA Assay

$390.00

The Human C3 ELISA Assay Kit is a highly sensitive two-site enzyme linked immunoassay (ELISA) for measuring C3 in biological samples of humans.

SKU: HC331-K01 Categories: , ,

Human C3 ELISA Assay

The Human C3 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 2.524 ng/ml
Standard Range: 12.5 – 400 ng/ml
Incubation Time: 60 minutes
Sample Type: Plasma, Serum
Sample Size: 100 µl


Assay Principle

The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the C3 present in samples reacts with the anti-C3 antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing, anti-C3 antibodies conjugated with horseradish peroxidase (HRP), are added. These enzyme-labeled antibodies form complexes with the previously bound C3. Following another washing step, the enzyme bound to the immunosorbent is assayed by the addition of a chromogenic substrate, 3,3’,5,5’-tetramethylbenzidine (TMB). The quantity of bound enzyme varies directly with the concentration of C3 in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of complement C3 in the test sample. The quantity of complement C3 in the test sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution.


Specimen Collection and Handling
Blood should be collected by venipuncture. The serum should be separated from the cells after clot formation by centrifugation. For plasma samples, blood should be collected into a container with an anticoagulant and then centrifuged. Care should be taken to minimize hemolysis, excessive hemolysis can impact your results. Assay immediately or aliquot and store samples at -20°C. Avoid repeated freeze thaw cycles.


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Additional Information

Assay Background


A number of serum proteins participate in acute inflammatory reactions. These include the complement, coagulation and kinin systems as well as a number of other proteins, known as acute phase proteins, that regulate acute inflammation. The complement system is a complex set of up to 20 serum proteins. The most abundant and pivotal of the complement components is C3, which has a molecular weight of about 187 kD and consists of an alpha and beta chain.

Typical Standard Curve


Manual

Product Manual