ß-Galactosidase Detection Reagents Selection Guide
X-Gal (5-bromo-4-chloro-3-indolyl-ß-D-galactopyranoside) is widely used as a chromogenic substrate for ß-galactosidase in the reporter gene assay. X-Gal is a small molecule which consists of Galactose moiety and 5-bromo-4-chloro-3-indole moiety. When X-Gal is cleaved, an insoluble blue precipitate is formed.
Fluorescein Di-ß-D-galactopyranoside (FDG), which is a fluorescein-based galactosidase substrates, and is used for detection of ß-galactosidase. This substrate forms a soluble green fluorescent product upon hydrolysis.
Although FDG enters viable cells, its permeability is insufficient for the use of live cell imaging. C12FDG, which is a variant of FDG, has been modified to have higher cell permeability. C12FDG can be loaded in the live cells; however, high concentration and long incubation time are required for loading into live cells.
To overcome these issues, Dojindo has introduced SPiDER-ßGal. By the enzymatic reaction, SPiDER-ßGal produces a quinone methide and forms covalent bond with nucleophic functional groups of the proteins in the vicinity of the molecules, producing a fluorescent product that is well retained by cells. Unlike commercially available substrates, SPiDER-ßGal allows a single-cell analysis under the lowest level of substrate concentration.
ß-Galactosidase Detection Reagents Selection Guide
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Product Name
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SPiDER-ßGal
Read More
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FDG
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C12FDG
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X-Gal
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ONPG
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Measurement
|
Fluorescence
|
Fluorescence
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Fluorescence
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Colorimetric
|
Colorimetric
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Wavelength
(ex/em)
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493,525/560
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490/514
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497/518
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615 nm
|
420 nm
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Detection Method
|
Fluorescence
microscope
Flow cytometry
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Fluorescence
microscope
Flow cytometry
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Fluorescence
microscope
Flow cytometry
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Microscope
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Absorbance
microplate reader
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For Live Cell
|
○ |
×
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△
|
×
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×
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For Fixed Cell
|
○ |
△
|
△
|
○
|
△
|
Intracelluler Retention
|
◎
|
×
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△
|
○
|
×
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Final Conc. of Cell Staining
|
1 µmol/l
|
100 ~ 2 mmol/l
|
33 µmol/l
|
2.5 mmol/l
|
2.3 mmol/l~*
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Incubation Time of Cell Staining
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15 min.
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20-25 min.
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20-60 min.
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Overnight
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N/A
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Number of Enzyme Reactive Site
|
1
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2
|
2
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1
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1
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Product Code
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SG02-10
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-
|
-
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-
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-
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* This Conc. is for ELISA format.