Human C5a ELISA Kit


The Eagle Biosciences Human C5a ELISA kit is to be used for the quantitative determination of human C5a/C5a-desArg in serum and plasma. The human C5a ELISA kit is not intended for measuring recombinant human C5a/C5adesArg. The Eagle Biosciences Human C5a ELISA Kit is for research use only and not for diagnostic or therapeutic procedures.

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Human C5a ELISA Kit

The Human C5a ELISA Kit is For Research Use Only

Sizes: 1×96 wells and 2×96 wells
Sensitivity: 0.3 ng/ml
Standard Range: 0.3 to 20 ng/ml
Incubation Time: 3.5 hours
Sample Type: Serum and plasma
Sample Size: 100 µl
Alternative Names: Complement Protein 5

Assay Background

The complement system is an important factor in innate immunity. During complement activation of complement protein 5 (C5), C5 is proteolytically cleaved and the anaphylatoxic peptide C5a is generated. C5a is a small polypeptide consisting of 74 aminoacids (11kDa) and is derived from the N-terminus of the alpha chain. C5a itself is very short-lived and in serum is cleaved rapidly into the more stable, though biologically still active C5a-desArg. Therefore, measurement of C5a-desArg allows reliable conclusions about the level of complement activation in the test samples. For convenience, both forms will be referred to in the following text as C5a.
C5a acts as a potent anaphylatoxin causing smooth muscle contraction, vasodilatation, increased vascular permeability, basophil and mast cell degranulation and lysosomal enzyme release. In addition, C5a is a potent chemotactic factor for eosinophils, basophils and monocytes. C5a is involved in inflammatory reactions seen in gram-negative bacterial sepsis, trauma, ischemic heart disease, post-dialysis syndrome and a variety of autoimmune diseases. Elevation of C5a is associated with increased cardiovascular risk in patients with advanced atherosclerosis. Also, C5a is closely associated with the capillary leak syndrome in leukemic children after bone marrow transplantation. C5a is also a marker in urine for predicting the onset of acute graft rejection after kidney transplantation. Activation products of the complement cascade contain neo-epitopes that are not present in the individual native components. The human C5a ELISA kit is based on a catching monoclonal antibody that recognizes a neo-epitope on C5a-desArg. This prevents cross-reactivity with C5.

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Product manufactured in the U.S.A.

Additional Information

Assay Principle

The human C5a ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 3½ hours. The efficient format of 2 plates 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 recognizing human C5a. Biotinylated tracer antibody will bind to the captured human C5a. Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody. Streptavidin-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. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the human C5a standards (log). The human C5a concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.


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