β -galactosidase,
a widely used reporter enzyme, is used in the study of gene expression, protein-protein
interactions,1 and normalization of transfection efficiency.2 β-galactosidase,
enzyme encoded by the lacZ gene of E. coli, catalyzes the hydrolysis
of β-galactosides into monosaccharides.
AnaSpec is
pleased to announce the release of 3 new assay kits for measuring β-galactosidase
activity – the SensoLyte®
MUG β-Galactosidase Assay Kit (Ex/Em= 360/460 nm), SensoLyte®
FDG β-Galactosidase Assay Kit (Ex/Em= 490/520 nm), and SensoLyte®
ONPG β-Galactosidase Assay Kit (Abs=420 nm). One advantage of using
a fluorogenic substrate, as is the case in the first two assays, is that it
is possible to monitor expression in individual cells as well as analyze the
heterogeneity of expression in cell populations.
3 Another major advantage is that fluorimetric
assays are several times more sensitive than colorimetric assays (see Table
1).
Table 1.
Comparison of the 3 β–Galactosidase assay kits
| |
SensoLyte®
MUG
β-Galactosidase Assay
|
SensoLyte®
FDG
β-Galactosidase
Assay
|
SensoLyte®
ONPG
β-Galactosidase
Assay
|
| Read-out |
Fluorimetric |
Fluorimetric |
Colorimetric |
| Wavelengths |
Ex/Em= 360/460 nm |
Ex/Em= 490/520 nm |
Abs=420 nm |
| Substrate Used |
4-Methylumbelliferyl β-D-galactopyranoside
(MUG) |
Fluorescein di-b-D galactopyranoside (FDG) |
o-Nitrophenyl β-D-galactopyranoside
(ONPG) |
| Hydrolysis Product |
4-Methylumbelliferone (4MU) |
Fluorescein |
o-Nitrophenol |
| Lowest Detection Limit |
15 fg |
10 pg |
100 pg |
| Cell Lysis Requirement |
With or without cell lysis3 |
With or without cell lysis |
Cell lysis required |

Figure 1. Detection of β-galactosidase
activity with the SensoLyte® MUG β-Galactosidase Activity Assay Kit (A),
SensoLyte® FDG β-Galactosidase Activity Assay Kit (B), and SensoLyte® ONPG
β-Galactosidase Activity Assay Kit (C); with lowest detection limits at
15 fg, 10 pg and 0.1 ng, respectively.
References
1. Rossi, F. et al. PNAS 94, 8405 (1997).
2. Thompson CD et al. Biotechniques
27, 824 (1999).
3. Plovins, A. et
al. Appl. Environ. Microbiol. 60, 4638 (1994).
|