The monoclonal antibody 2991 recognizes the neo-epitope of human C3a/C3a-desArg that is formed upon cleavage of the complement protein C3 into C3a and C3b. The complement system is an important part of the humoral response in innate immunity, consisting of three different pathways. The third complement component, C3, is central to the classical, alternative and lectin pathways of complement activation. Activation products of the complement cascade contain neo-epitopes that are not present in the individual native components. Monoclonal antibodies detecting neo-epitopes have been used for direct quantification of activation at different steps in the complement cascade. The synthesis of C3 is tissue-specific and is modulated in response to a variety of stimulatory agents. C3 is the most abundant protein of the complement system with serum protein levels of about 1.3 mg/ml. An inherited deficiency of C3 predisposes the person to frequent assaults of bacterial infections. In ulcerative colitis, and idiopathic chronic inflammatory bowel disease, the deposition of C3 in the diseased mucosa has been reported. Proteolysis by certain enzymes results in the cleavage of C3 releasing C3a anaphylatoxin and C3b. C3a is a protein of 74 amino acids. C3a itself is very short-lived and in serum is cleaved rapidly into the more stable C3a-desArg (also called acylation stimulating protein, ASP) . Therefore, measurement of C3a-desArg allows reliable conclusions about the level of complement activation in the test samples. C3a is a mediator of local inflammatory processes. It induces smooth muscle contraction, increases vascular permeability, and causes histamine release from mast cells and basophilic leukocytes. C3a is involved in inflammatory reactions seen in gram-negative bacterial sepsis, trauma, ischemic heart disease, post-dialysis syndrome and a variety of autoimmune diseases. Normal values in plasma of control persons range between 48 150 ng/ml (median: 86.4 ng/ml, SD: 29.3 ng/ml)
Monoclonal antibody 2991 recognizes both C3a and C3a-desArg, with at least a 5x higher preference for C3a-desArg.
Immuno assays, Western blot
W: A reduced sample treatment and SDS-PAGE was used. The band size is ~8 kDa.
Dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50.
Activated human serum or purified human C3a desArg protein
For Western blot, any irrelevant protein can be used.