ELISA, Immunohistochemistry, Western Blot. Anti-Beta Galactosidase is suitable for immunoblotting (western or dot blot), ELISA, immunofluorescence microscopy, immunoprecipitation, conjugation and most immunological methods requiring high titer and specificity. The antibody recognizes both frozen tissue sections, paraffin embedded tissue and 4% paraformaldehyde fixed tissue for most immunohistochemical analysis. A 1:5,000 dilution has been reported to be successful for staining by immunoblot of beta-galactosidase fusion proteins after transfer using a semi-dry transfer apparatus. A 1:1,500 dilution has been reported to detect beta-galactosidase in adult rat spinal cord tissue after infection with helper-dependent adenovirus expressing lacZ. In this particular experiment, tissue was perfused with 4% paraformaldehyde and cryostat-cut (35 μm) to produce free-floating sections. A 1:5,000 dilution has been reported to be successful for staining of brain sections from transgenic mice expressing nuclear beta-galactosidase when assayed by immunofluorescence microscopy. A 1:5,000 dilution has been reported for immunofluorescent staining of methanol fixed, devitellinized Drosophila embryos. Although a wide range of conditions was reported to be effective, a 1:10,000 dilution was noted to show no background and to be suitable for double labeling experiments. Optimal titers for other applications should be determined by the researcher.
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Description
Beta Galactosidase is an IgG fraction antibody purified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Peroxidase, and anti-Rabbit Serum as well as purified and partially purified Beta Galactosidase [E.coli]. Cross reactivity against Beta Galactosidase from other tissues and species may occur but have not been specifically determined. Very low background staining has been reported in various assays.