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About This Item
Empirical Formula (Hill Notation):
C27H28N6O · 3HCl · xH2O
CAS Number:
Molecular Weight:
561.93 (anhydrous basis)
NACRES:
NA.47
PubChem Substance ID:
UNSPSC Code:
12171500
MDL number:
Beilstein/REAXYS Number:
1234011
Quality Level
assay
≥98% (HPLC)
form
powder
technique(s)
titration: suitable
color
yellow
pH
1.7 (20 °C)
solubility
H2O: 20 mg/mL, phosphate buffer: precipitates
suitability
suitable for fluorescence
application(s)
diagnostic assay manufacturing
hematology
histology
storage temp.
−20°C
SMILES string
Cl[H].Cl[H].Cl[H].CCOc1ccc(cc1)C2=NCc3cc(ccc3N2)C4=NCc5cc(ccc5N4)N6CCN(C)CC6
InChI
1S/C29H32N6O.3ClH/c1-3-36-25-8-4-20(5-9-25)28-30-18-22-16-21(6-10-26(22)32-28)29-31-19-23-17-24(7-11-27(23)33-29)35-14-12-34(2)13-15-35;;;/h4-11,16-17H,3,12-15,18-19H2,1-2H3,(H,30,32)(H,31,33);3*1H
InChI key
FYEVKHPLBHLWHK-UHFFFAOYSA-N
General description
Fluorescent properties of bisBenzimide H 33342:
Free dye: Excitation maximum = 340 nm, Emission maximum = 510 nm (5 mM HEPES, 10 mM NaCl, pH 7.0) DNA complex: Excitation maximum = 355 nm, Emission maximum = 465 nm (5 mM HEPES, 10 mM NaCl, pH 7.0)
Free dye: Excitation maximum = 340 nm, Emission maximum = 510 nm (5 mM HEPES, 10 mM NaCl, pH 7.0) DNA complex: Excitation maximum = 355 nm, Emission maximum = 465 nm (5 mM HEPES, 10 mM NaCl, pH 7.0)
Application
Bisbenzimide Hoechst 33342 is a specific stain for AT-rich regions of double-stranded DNA and has been shown to displace several known DNA intercalators. This fluorescent dye has been used in sorting living cells based on DNA content, used in flow cytometry for the determination of DNA content, and for the visualization of chromatin distribution in living cells. It has been used to detect BrdU incorporation into cells and in studying the initial stages apoptosis and cellcycle distribution., Chromosomes that are dividing or replicating will not stain with this dye.
Useful for staining DNA, chromosomes and nuclei. May be used for fluorescence microscopy or flow cytometry.
Excitation max. = 346 nm
Emission max. = 460 nm
Excitation max. = 346 nm
Emission max. = 460 nm
Biochem/physiol Actions
Membrane-permeable, fluorescent DNA stains with low cytotoxicity that intercalate in A-T regions of DNA.
Preparation Note
This product is soluble in water (50 mg/ml), yielding a clear solution. The pH of a 2% solution is 1.9. It has been observed that this material will precipitate from phosphate buffer solutions.
Aqueous solutions are stable for 1 month if kept in the dark at 2-8 °C.
Aqueous solutions are stable for 1 month if kept in the dark at 2-8 °C.
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Storage Class
11 - Combustible Solids
flash_point_f
Not applicable
flash_point_c
Not applicable
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M G Ormerod et al.
Cytometry, 14(6), 595-602 (1993-01-01)
We have recently developed a method for the separation and quantification of viable apoptotic cells without the need for permeabilisation or fixation of the cells. The method is based on the observation that apoptotic rat thymocytes fluoresce more brightly than
M Gregoire et al.
Experimental cell research, 152(1), 38-46 (1984-05-01)
Chromatin distribution was visualized in living cells with the selective DNA fluorochrome Hoechst 33342. This dye was shown to be non-toxic on the rat kangaroo PTO cell line by measuring the labelled cell growth rate. The aim of this work
The transforming activity of Wnt effectors correlates with their ability to induce the accumulation of mammary progenitor cells.
Liu BY, et al.
Proceedings of the National Academy of Sciences of the USA, 101, 4158-4163 (2004)