Additional Text

Concentration

1 mg/ml

Short Description

Tetramethylrhodamine isothiocyanate-conjugated IgG fraction of polyclonal rabbit antiserum to Bovine IgG, heavy and light chains. To identify and measure IgG, antigen or antibody, at the cellular and subcellular level by immunofluorescence staining of appropriately treated cell and tissue substrates. If two Biorbyt immunoconjugates with different labels but originating from the same host animal species are used for direct two-color staining, they can safely be mixed and used in a single step, without the danger of interaction between the two reagents. 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 1:20 and 1: 80.

Note

For research use only.

Antibody Clonality

Polyclonal

Application Notes

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 antiserum has been tested in double radial immunodiffusion with the following results: cat + chicken - dog + duck - goat ++ guinea pig + hamster + horse ++ human ++ mouse + monkey + pigeon - rat + sheep ++ swine + turkey - A negative cross-reaction in double radial immunodiffusion does not exclude some reaction in more sensitive techniques.Conjugation Notes: A proprietary technique for the binding to TRITC is used, followed by several purification steps to remove free reactants and protein aggregates. After each step activity and specificity are tested in a variety of techniques. The conjugate is lyophilized to assure stability and long shelf life. Tetramethylrhodamine isothiocyanate isomer R. It has an orange-red fluorescence. Excitation: 554 nm, emission: 573 nm. To avoid nonspecific background staining, specially synthesized and exceptionally pure crystalline isomer R has been used instead of the usual racemic mixture. Although its fluorescence efficiency is less than of FITC, TRITC conjugates have the advantage of significantly less photo bleaching. This facilitates their use in quantitative cell-counting procedures.Experiment Notes: Hyperimmune antisera with strong precipitating activity are selected for fractionation by salt-precipitation and purification of the IgG fraction by DEAE-chromatography.

Storage Note

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