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. of this antiserum has not been tested in detail.
Characteristics
Fluorescein isothiocyanate-conjugated IgG fraction of polyclonal goat antiserum to C5 fragment of human complement
Purification
Adsorption: Immunoaffinity adsorbed using insolubilized antigens as required, to eliminate antibodies cross-reacting with other with other plasma proteins. The use of insolubilized adsorption antigens prevents the presence of excess adsorbent protein or immune complexes in the antiserum. The IgG (7S) fraction is isolated and purified from the antiserum and contains the bulk of the defined antibody specificity. It is free of other serum proteins as tested by immunoelectrophoresis and double radial immunodiffusion.
Immunogen
C5 is a component of the complement system. It has a molecular weight of 190,000. The activated protein forms with the complement factors C6-C9 the membrane attack-complex. C5 is isolated and purified from pooled normal human serum. Freund’s complete adjuvant is used in the first step of the immunization procedure.
C5
Reactivity: Human
WB, IA
Host: Mouse
Monoclonal
568
unconjugated
Application Notes
The fluorescent immunoconjugate to human C5 is used to determine the presence and pattern using immunohistochemical staining techniques. Determinations of individual C components can be very useful in defining the exact location of a defect. 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 are usually between 1:10 and 1:40.
Restrictions
For Research Use only
Format
Lyophilized
Concentration
IgG protein concentration 10 mg/ml. Fluorescein/IgG protein molar ratio (F/P) approximately 1.4. No foreign proteins added.
Buffer
Fluorochrome-coupled purified hyperimmune IgG lyophilized from a solution in phosphate buffered saline (PBS, pH 7.2)
Preservative
Without preservative
Storage
4 °C/-20 °C
Storage Comment
The lyophilized conjugate is shipped at ambient temperature and may be stored at +4°C, prolonge d storage at or below -20°C. It is reconstituted by adding 1 ml sterile di stilled 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 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.
Tested in immunoelectrophoresis and double radial immunodiffusion against normal human serum, a single precipitin line is obtained which gives a reaction of full identity with the isolated immunogen. The product does not react with any other proteins component of human serum or plasma