The antiserum is reacting with to the Fab subunits of the IgG, IgA, IgM and other Ig classes of both light chain type, with their Fab or F(abÕ)2 subunits and with free immunoglobulin light chains of kappa and lambda type. It does not react with any non-Ig protein in human serum, as tested by immunoelectrophoresis and double radial immunodiffusion. Direct staining of fixed cell and tissue substrates, to demonstrate the intracellular presence of free of Ig bound subunits of both kappa and lambda type. The absence of the Fc domain in the conjugate ensures minimal interaction with the tissue components and cell surfaces other than the primary antibody activity. This conjugate is primarily intended for use in cell surface membrane staining procedures, to identify and quantitate Ig on B cells, especially if interference by Fc activity is expected.. 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.
Tetramethylrhodamine isothiocyanate-conjugated F(abÕ)2 fragments of polyclonal Goat IgG antibodies to Human Fab IgG
ELISA,Immunocytochemistry,Immunohistochemistry (frozen),(In)direct immunofluorescence
Fields of Interest
Purified Fab prepared from normal IgG isolated from pooled human serum. FreundÕs complete adjuvant is used in the first step of the immunization procedure.
TRITC-coupled goat F(abÕ)2 antibodies lyophilized from a solution in phosphate buffered saline (PBS, pH 7.2). No preservative added.
Inter-species cross-reactivity is a normal feature of antibodies to immunoglobulins, since Ig of different species frequently share antigenic determinants. Cross-reactivity of this conjugate has not been tested in detail.
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 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. Lyophilized at +4¡ C--at least 10 years reconstituted at or below -20¡ C--3-5 years reconstituted at +4¡ C--7 days