This antibody detects 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. Immunoelectrophoresis gives a single precipitin arc against anti-rabbit serum as well as purified and partially purified Beta-galactosidase [E.coli].
Purification
Multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer
Immunogen
Full length native Beta-galactosidase isolated from E.coli
Suitable for Immunoblotting (Western or dot blot), ELISA, Immunofluorescence microscopy, Immunoprecipitation, conjugation and most immunological methods requiring high titer and specificity. Western blot : 1/5,000-1/10,000. 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). ELISA : 1/10,000. Immunohistochemistry : 1/1,500. The antibody recognizes both Frozen tissue sections, Paraffin embedded tissue and 4 % paraformaldehyde fixed tissue for most immunohistochemical analysis: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 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). A 1/5,000 dilution has been reported to be successful for staining brain sections from transgenic mice expressing nuclear Beta-galactosidase when assayed by Immunofluorescence microscopy .
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
4 °C,-20 °C
Storage Comment
Store lyophilized at 2-8°C for 6 months or at -20°C long term. After reconstitution store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C long term. Dilute only prior to immediate use. Avoid repeated freezing and thawing. Shelf life: one year from despatch.
Beta-galactosidase is coded by a gene (lac z) in the lac operon of Escherichia coli. It is a metalloenzyme that splits lactose into glucose and galactose. It hydrolyzes terminal, non-reducing beta-D-galactose residues in beta-D-galactosides. Activation by cations seems to be substrate dependent. K + , Na , NH 4 , Rb , Cs+ and Mn ++ all activate enzyme activity based upon the substrate used. Anti Beta-galactosidase antibody recognizes the enzyme beta-galactosidase, or ß-galactosidase, that is a component of assays used frequently in genetics, molecular biology (see X-gal) for a blue white screen, and other life sciences. IPTG induces production of ß-galactosidase by binding and inhibiting the lac repressor. Since it is highly expressed and accumulated in lysosomes in senescent cells, it is used as a senescence biomarker both in vivo and in vitro in qualitative and quantitative assays, despite its limitations. Anti Beta-galactosidase antibody is ideal for investigators involved in enzyme research.