In immunoelectrophoresis against fresh rat 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 rat serum or plasma. The biotin immunoconjugate to rat 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. In ELISA and Western blotting to identify rat C3c in serum or other body fluids. As a second step an avidin or streptavidin conjugate of the user’s choice has to be used. 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 dilutions for histochemical and cytochemical use are usually between 1:100 and 1:500; In ELISA and comparable non-precipitating antibody-binding assays between 1:1,000 and 1:5,000.
Antibody come from
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 rat serum. Freund’s complete adjuvant is used in the first step of the immunization procedure.
Biotin-coupled purified hyperimmune IgG lyophilized from a solution in phosphate buffered saline (PBS, pH 7.2) No preservative added, as it may interfere with the antibody activity.
Antigen-antibody binding interaction
Goat anti Rat C3c antibody conjugated with Biotin Antibody
Antibody's reacts with these species
The antiSerum does not cross-react with any other component of Rat plasma. Inter-species cross-reactivity is a normal feature of antibodies to plasma proteins since they frequently share antigenic determinants. Crossreactivity of this antiSerum has not been tested in detail.
Biotin-conjugated IgG fraction of polyclonal Goat antiSerum to C3c fragment of Rat complement factor C3.
ELISA,Immunocytochemistry,Immunohistochemistry (paraffin),Dot blot,Immunoblotting
Antibody's suited for
ELISA,Immunocytochemistry,Immunohistochemistry (paraffin),Dot blot,Immunoblotting.
The lyophilized conjugate is shipped at ambient temperature and may be stored at +4°C; prolonged storage at or below -20°C. It is reconstituted by adding 1 ml sterile distilled water, spun down to remove insoluble particles, divided into small aliquots, frozen and stored at or below -20°C. Prior to use, an aliquot is thawed slowly 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. Lyophilized at +4° C--at least 10 years. Reconstituted at or below -20° C--3-5 years. Reconstituted at +4° C--7 days
This antibody needs to be stored at + 4°C in a fridge short term in a concentrated dilution. Freeze thaw will destroy a percentage in every cycle and should be avoided.
If you buy Antibodies supplied by nordc they should be stored frozen at - 24°C for long term storage and for short term at + 5°C.Biotin conjugates can be detected by horseradish peroxidase, alkaline phosphatase substrates or anti biotin conjugated antibodies. Avidin and Streptavidin bind to the small biotin and are couple to HRP or AP for ELISA. To break the streptavidin Biotin bond we suggest to use a 6 molar guanidine HCl solution with acidity of pH 1.6.
Capra aegagrus hircus,Rattus norvegicus
Rats are used to make rat monoclonal anti mouse antibodies. There are less rat- than mouse clones however. Rats genes from rodents of the genus Rattus norvegicus are often studied in vivo as a model of human genes in Sprague-Dawley or Wistar rats.