The dye is nonfluorescent (and quite mutagenic, I believe).
Anal Biochem 1985 Aug 15;149(1):117-20
Related Articles, Books
Sensitive assay systems for detection of hemoglobin with
2,7-diaminofluorene: histochemistry and
colorimetry for erythrodifferentiation.
Kaiho S, Mizuno K.
Sensitive and rapid assays, colorimetry and histochemistry,
for hemoglobin in erythroid cells are established. The assays are based on
pseudoperoxidase activity of hemoglobin using
2,7-diaminofluorene as a hydrogen donor for the peroxidase, instead of benzidine
which is widely benzidine which is widely used for the
detection of small amounts of hemoglobin but which is a potent carcinogen and
has been banned from laboratory use. In the presence of
hydrogen peroxide, hemoglobin catalyzes the formation of a blue compound
(fluorene blue), which has a broad absorption band between 500
and 690 nm with a peak at 610 nm, from 2,7-diaminofluorene. The
reagent is safe to use in the laboratory. The methods could be
applied to the detection of hemoglobin in Friend erythroleukemia cells
induced to cell differentiation along the erythroid pathway by
dimethyl sulfoxide.
Geoffrey Osborne wrote:
> Hi,
> Just wondering if anyone has experience assessing the specific oxidation
> of 2,7-diaminofluorene (DAF) by pseudoperoxidase activity of hemoglobin
> (Hb)to generate fluorene blue (FB) by flow cytometry?
> There seems to numerous plate reader based assays i.e. colourimetric, but
> I haven't been able to find out if the fluorene blue is at all fluorescent.
> Any advice appreciated.
> Thanks
>
> Geoffrey Osborne
>
> Specialist, Flow Cytometry,
> John Curtin School of Medical Research,
> The Australian National University,
> Canberra, 0200, ACT. AUSTRALIA
> email: geoff.osborne@anu.edu.au
> http://jcsmr.anu.edu.au/facslab/facshome.html
>
> (61 2) 6125 3694.
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