The gene of beta-galactosidase from E. coli is widely used as a reporter gene assay marker. Although X-gal is well known reagent to detect beta-galactosidase in cell or tissue samples, the assay using these reagents require to fix cells or tissues due to the poor cell-permeability. In addition, so far developed the assay using fluorescence reagents can not clearly differentiate beta-galactosidase-expressed cells or regions.To overcome these issues, Urano, Kamiya and co-workers have successfully developed a stain ideally possesses cell-permeability and the ability to retain in intracellular region.1)By the enzymatic reaction, the beta-Galactosidase stain immediately forms a quinone methide that acts as electrophile when proteins containing nucleophilic functional groups nearby the molecules. By the probe undergoes the reaction with a protein, the conjugates become fluorescent compounds. Thus, it allows a single-cell analysis because it does self-immobilizing to the intracellular proteins.Exciation Maximum: 530 nm (+/- 10 nm)Emission Maximum: 550 nm (+/- 10 nm)Reference: 1) Y. Urano, M. Kamiya, T. Doura, WO 2015174460, A1, (19, November, 2015). Find more infomation here 94433
Synonyms: 6'-(Diethylamino)-4'-(fluoromethyl)spiro[isobenzofuran-1(3H),9'-[9H]xanthen]-3'-yl beta-D-galactopyranoside; SPiDER-betaGal; SPiDER-beta Gal; (2S,3R,4S,5R,6R)-2-{[3'-(Diethylamino)-5'-(fluoromethyl)-3H-spiro(isobenzofuran-1,9'-xanthen)-6'-yl]oxy}-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol
Molecular Formula: C31H34FNO8
Molecular Weight: 567.6
Linear Structural Formula: C31H34FNO8
Storage: 2-8C
Application: Preparation of 1 mmol stain DMSO stock solutionAdd 35 mul of DMSO to a tube of stain solution (20 mug) and dissolve it with pipetting.*Store the stock solution at -20C.Preparation of 1 mumol/l stain working solutionDilute the stain DMSO stock solution with Hanks' HEPES buffer to prepare 1 mumol/l stain working solution.*Hanks' HEPES buffer is recommended to maintain cell condition.General protocol:staining1. Prepare cells for the assay. 2. Discard the culture medium and wash the cells with Hanks' HEPES buffer twice. 3. Add an appropriate volume of stain working solution.4. Incubate at 37oC for 15 minutes.5. Observe the cells under a fluorescence microscope or by a flow cytometer.*After staining, the cells can be observed even without washing. However, you can wash it as needed.