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CELL CULTURE & TRANSFECTION 1
Reporter Detection Kits
Gene reporter systems are extensively used for the study of eukaryotic gene expression and regulation. Although
they play a significant role in numerous applications, they most frequently serve as indicators of transcriptional
activity in cells. The reporter gene of choice is combined to a promoter sequence and can be subsequently assayed
in vitro or in vivo. An ideal reporter gene is not endogenously expressed by the target cells creating background
signals and can be detected through assays that are easy, sensitive and reliable.
InvivoGen provides two reporter gene systems that can be measured by histochemical staining of cells or tissues:
• b-Galactosidase (LacZ) reporter gene system
• Secreted embryonic alkaline phosphatase (SEAP) reporter gene system
Furthermore, InvivoGen has developed two specific media that allow fast and convenient detection of secreted
AP, called QUANTI-Blue and HEK-Blue Detection (see pages 14-15).
LacZ Reporter Gene System
The E. coli lacZ gene encoding b-galactosidase is the classical histochemical reporter gene. b-Galactosidase catalyzes the hydrolysis of X-Gal
producing a blue precipitate that can be easily visualized under a microscope. The LacZ Staining Kits provide simple and convenient methods
for the visual detection of LacZ expression within cells or tissues.
LacZ Reporter Gene
InvivoGen provides the LacZDCpG gene, a humanized and CpG-free
allele of the LacZ gene. This CpG-free gene is ten times more active than
the wild-type gene in mammalian cells. It can be used for in vitro or in
vivo applications. The LacZDCpG gene is provided in the pSELECT-zeo-
LacZ plasmid under the control of the EF-1a/HTLV composite promoter.
The pSELECT-zeo-LacZ plasmid is selectable with Zeocin in mammalian
cells and in bacteria.
Contents and Storage
The LacZDCpG gene is provided as 20 μg of lyophilized DNA with 4
pouches of E. coli Fast-Media ® Zeo (2 TB and 2 Agar, see p45). Plasmid is
shipped at room temperature. Store at -20°C for up to one year.
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LacZ Staining Kits
www.invivogen.com/reporter-detection
InvivoGen provides two LacZ staining kits:
- LacZ Cell Staining Kit allows you to determine the percentage of
transfected cells expressing the lacZ gene. All reagents are provided to
prepare the fixative, staining and rinse solutions. The assay can be
completed in 30 minutes, and the blue precipitate can take from 30
minutes to overnight to appear. The LacZ Cell Staining Kit contains
sufficient reagent to stain one hundred 35 mm plates.
- LacZ Tissue Staining Kit allows the detection of transduced cells
expressing the lacZ gene within fresh or frozen tissues. All reagents are
provided to prepare the buffer, fixative and staining solutions. The staining
of the tissues can be observed in 5 to 24 hours. The LacZ Tissue Staining
Kit contains sufficient reagent to prepare 100 ml solutions.
Contents and Storage
The LacZ Cell Staining Kit contains: 10X PBS, X-Gal, Potassium
Ferrocyanide, Potassium Ferricyanide, MgCl2 and 10X Fixative Solution.
The LacZ Tissue Staining Kit contains: 10X PBS, X-Gal, Potassium
Ferrocyanide, Potassium Ferricyanide, MgCl2, Glutaraldehyde, Igepal and
Na deoxycholate.
Products are shipped at room teperature. Store at 4°C or -20°C
according to the product label. All reagents are stable for 6 months.
PRODUCT QUANTITY CAT. CODE
pSELECT-zeo-LacZ 20 μg psetz-lacz
LacZ Cell Staining Kit 1 kit rep-lz-c
LacZ Tissue Staining Kit 1 kit rep-lz-t