Background
The antibody identifies the inactivated C3c-component and is useful for detection of C3-deposit in tissues after complement activation. Of special interest is the analysis of C3c in different kinds of pemphigus. In this disease deposits of C3 are mainly found in the basal membrane of the skin and to some extend in the basal keratinocytes of the epidermis. C3c-component.

Source
C3c is the major fragment resulting from C3 cleavage by C3 convertase and factor I. It is composed of an intact beta chain bound to two fragments of the alpha chain. C3c is isolated and purified from pooled normal human serum. Freund’s complete adjuvant is used in the first step of the immunization procedure.

Immunogen: Purified Complement 3 from human serum

Product
Ion-exchanged purified IgG conjugated with Fluorescein Isothiocyanate Isomer 1 in PBS pH 7.2; contains sodium azide (0,09%)**

Purification Method: Ion-exchanged purified IgG conjugated with Fluorescein Isothiocyanate Isomer 1 in PBS pH 7.2; contains sodium azide (0,09%)**

Specificity
The antiSerum does not cross-react with any other component of Human plasma. Inter-species cross-reactivity is a normal feature of antibodies to plasma proteins since they frequently share antigenic determinants. Cross-reactivity of this antiSerum has not been tested in detail.

Species Reactivity: Human

Applications
IHC (F); The dilution has to be evaluated individually for optimal results. In immunoelectrophoresis against fresh human serum, a single precipitin line is obtained in the beta-1 region representing native C3. Against serum containing partly activated C3, a precipitin line is obtained which extends from the beta-1 into the alpha-2 region, demonstrating a gradient. In old serum containing totally activated C3 a single precipitin line in the alpha-2 region is obtained. Antisera to C3c cab also react with the fragments C3b, C3bi and smaller fragments, since they all carry antigenic determinants of the C3c domain. The product does not react with any other proteins component of human serum or plasma. The fluorescent immunoconjugate to human C3c is used to determine the presence and pattern of C3 in tissue lesions using immunohistochemical staining techniques. Locally deposited immune complexes in tissue usually contain complement, pointing to activation of the classical pathway. Complement activation in vivo implies active disease and may contribute to the elicitation of the pathogenesis and he extent of tissue destruction. Sometimes the diagnosis can be based on directly on laboratory findings. This immunoconjugate is not pre-diluted. The optimum working dilution of each conjugate should be established by titration before being used. Excess labelled antibody must be avoided because it may cause high unspecific background staining and interfere with the specific signal.

Working Concentration: The dilution of the antibody has to be titrated for the specific applications.

Positive Control: Skin (basal membrane) in bullous pemphigoid

Storage
2-8°C, protected from light. Prolonged storage at or below -20°C. Prior to use, an aliquot is thawed slowly in the dark at ambient temperature, spun down again and used to prepare working dilutions by adding sterile phosphate buffered saline (PBS, pH 7.2). Repeated thawing and freezing should be avoided. Working dilutions should be stored at +4°C, not refrozen, and preferably used the same day. If a slight precipitation occurs upon storage, this should be removed by centrifugation. It will not affect the performance of the immunoconjugate.

Shipping Conditions: The lyophilized conjugate is shipped at ambient temperature and may be stored at +4°C; prolonged storage at or below -20°C.

Caution
This product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals. It may contain hazardous ingredients. Please refer to the Safety Data Sheets (SDS) for additional information and proper handling procedures. Dispose product remainders according to local regulations.This datasheet is as accurate as reasonably achievable, but Exalpha Biologicals accepts no liability for any inaccuracies or omissions in this information.

References
1. Bernard P., Aucouturier P., Denis F., and J.M. Bonnetblanc (1990) Immunoblot analysis of IgG subclasses of circulating antibodies in bullous pemphigoid. Clin. Immunol. Immunopathol. 54(3); 484-494. 2. Feliciani C., Pour S.M., Toto P., Coscione G., and Amerio P. (1998) Direct immunofluorescence diagnosis of pemphigus without biopsy. J. Cutan. Med. Surg. 2(4); 209-211. 3. Buschman K.E., Seraly M., Thong H.Y., Deng J.S., Draviam R.P., and Abernet J.L. (2002) A predominant IgG4 subclass may be responsible for True-negative direct immunofluorescence in bullous pemphigoid. J. Cutan. Pathol. 29(5); 282-286.

Protein Reference(s)

Database Name: UniProt

Accession Number: P01024-CO3_HUMAN

Species Accession: Human

Safety Datasheet(s) for this product: