The E. coli gene lacZ encoding beta-galactosidase is an important reporter gene for detecting the expression of recombinant genes in animal cells. In situations where enzymatic detection of beta-galactosidase is not practical, anti-beta-galactosidase antibody has been widely used. Anti-beta-galactosidase antibody can also be used for purification of fusion proteins by immunoprecipitation or immunoadsorption. Furthermore, anti-beta-galactosidase antibody has been shown to be the preferred analytical tool for light microscopic evaluation of lacZ gene transfer, and the beta-galactosidase/anti-beta-galactosidase complex has been used as a model to assess the feasibility of a high-pressure immunodesorption process.
This antibody, raised in rabbit against E. coli beta-galactosidase, demonstrates high specificity for the enzyme and is suitable for immunological methods that require high titer and specificity such as immunoblotting (western or dot blot), ELISA, immunoprecipitation, and immunohistochemistry. The appropriate dilution of antibody for a particular application should be determined empirically. We suggest trying dilutions of about 1:5,000 to 1:50,000 for your initial experiments.
Beta-galactosidase is an exoglycosidase which hydrolyzes the beta-glycosidic bond formed between a galactose and its organic moiety. Deficiencies in the protein in humans can result in galactosialidosis or Morquio B syndrome. In E. coli, the gene of beta-galactosidase, the lacZ gene, is present as part of the inducible system lac operon, which is activated in the presence of lactose when glucose level is low. E. coli beta-galactosidase is commonly used in molecular biology as a reporter marker to monitor gene expression. Another popular use for beta-galactosidase is in blue/white screening to identify recombinant clones. Beta-galactosidase can be split into two peptides, lacZalpha and LacZOmega, neither of which is active by itself but when both are present together, spontaneously reassemble into a functional enzyme. This property is exploited in many cloning vectors. The presence or absence of an active beta-galactosidase may be detected through addition of artificial chromogenic substrates such as X-gal, fluorescent substrates such as Fluorescein di-beta-D-galactopyranoside (FDG), luminescent substrates, and others. Beta-galactosidase activity at pH 6 is an indicator of senescent cells not found in presenescent, quiescent or dividing cells.
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Protein Aliases: b-gal fusion protein; Beta-Gal; beta-gal fusion protein; betagal; bgal; ß-gal; ß-galactosidase; ßgal