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Immunofluorescent analysis of E. coli beta-Galactosidase (green) in pCMV-LacZ transfected HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a beta-Galactosidase monoclonal antibody (Product # MA1-152) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502). Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Bacteria|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Full length E. coli beta Galactosidase|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay dependent|
|Immunofluorescence (IF)||1:50 - 1:100|
|Western Blot (WB)||1:500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Western blot analysis of MA1-152 detects recombinant E. coli beta Galactosidase at ~110 kDa. This antibody also detects E. coli beta Galactosidase in pCMV-lacZ transfected HeLa cells.
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 in 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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Beta-Gal; GLB2; GSL; NGBE; PPCA; PPGB; RP3-337O18.1