Roti® -GelStain – Background and Safety Tests 99643-38-6
-GELSTAIN MOLECULAR DATA
Ready-to-use staining mixture prepared from a variety of reagents
Dye component: orange-red to brown powder
CAS [99643-38-6] Formula C21H28N4 MW: 336.47
DNA-binding properties: Benzimidazoles typically bind to the
minor groove of helical nucleic acid. No intercalation takes place
Application: Very well suited for staining of single and double stranded nucleic acids in agarose gels via addition
to the agarose (in-gel staining), to the running buffer, or via post-run staining. Less recommended for use via
addition to loading buffers. Not suitable for staining of PAA gels.
Light spectrum (bound to DNA):
Excitation maximum at 290-320 nm
Excitation also possible at 420 nm
Emission maximum at 515 nm
-GELSTAIN SAFETY PARAMETERS
Ethidium bromide is a widely used staining reagent in
nucleic acid electrophoresis and is putatively strong
carcinogenic. It has been always a great concern for
personnel and environment when using it in the
laboratories. In recent years, great efforts have been made to establish alternative nucleic staining dyes that are
safe to both, lab personnel and environment. Roti®-GelStain represents an alternative DNA/RNA staining dye that
addresses the laboratory and environmental safety concerns, and, additionally, makes life in electrophoresis labs
significantly more easy.
Mouse, subcutaneous: 250 mg/kg
Target organ: Blood
MAMMALIAN GENOTOXICITY ANALYSIS
AMES TEST SYSTEM
The test employed two Salmonella typhimurium strains, TA98 and TA1537, both of which carry mutation(s) in the
operon encoding for histidine biosynthesis. When these bacteria are exposed to mutagenic agents, under certain
conditions reverse mutation from amino acid (histidine) auxotrophy to prototrophy occurs, giving colonies of
revertants. Both strains of bacteria used in the assays are among those recommended by OECD 471 for use in
the Ames test. These two strains of S. typhimurium have been shown to be reliably and reproducibly responsive
between laboratories. In order to test the mutagenic toxicity of metabolized products, S9 fraction, a rat liver
extract, was used in the assays. The S9 fraction contains a mixture of several enzymes and is known to be able
to convert some chemicals into mutagens.
AMES TEST PROCEDURE
Roti®-GelStain along with ethidium bromide (EB) as a reference was tested under the same conditions.
DMSO was used for dissolving each dye to give the following stock concentrations:
0 (control), 1, 2.5, 5, 10, 25, 50, 75, 100, 250 and 500 µg/ml.
The following was added to each sterile culture tube containing 2.0 ml top agar: 0.1 ml of overnight cell culture
(TA98 or TA1537), 0.1 ml of each dye concentration for each dye or control chemical, and either 0.5 ml of
S9/Cofactor mix or 0.5 ml of phosphate buffered saline.
By using the above 10 stock solutions for each dye plus the control, the following per plate doses for each dye
were used: 0, 0.1, 0.25, 0.5, 1, 2.5, 5, 7.5, 10, 25, and 50 µg/ plate. These doses corresponded to a final dye
concentration of: 0, 0.04, 0.09, 0.19, 0.37, 0.93, 1.85, 2.78, 3.7, 9.3, and 18.5 µg/ml, respectively.
The contents of each tube were vortexed, poured onto Vogel-Bonner media plates, and evenly distributed. The
agar on the test plates was allowed to harden. The plates were inverted and incubated at 37 °C for 2 days.
Revertant colonies were counted using an automatic colony counter.
For strain TA1538, an increase in revertants of more than threefold over background indicates a positive result,
whereas an increase in revertants of more than two fold over background indicates a positive result for
mutagenicity in this test for strain TA98.
RESULTS OF AMES TESTS
Two test results are shown exemplarily.
At a dose of 1 µg/plate, ethidium bromide is significantly positive in Ames test (here approx. 19x and 23x above
background), whereas Roti®
-GelStain still is completely negative.
At a dose of 50 µg/plate, Roti®
-GelStain is just considered Ames test-positive (here appox. 4x and 8x above
background), while ethidium bromide is more or less beyond measures
更新时间：2015-06-15 15:48:08 【
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