Wash sequentially in 70% etoh containing 4% vv formaldehyde, 50% and 35% etoh, and 1x. Several related chemical compounds are used for similar purposes and are collectively called hoechst stains application. Hoechst 33342 staining dye solution ab228551 is a fluorescent stain for labeling dna in fluorescence microscopy. Hoechst 33342 is a popular cellpermeant nuclear counterstain that emits blue fluorescence when bound to dsdna. Bisbenzimide tends to bind to adeninethyminerich regions of dna and can decrease its density. Do not use other hoechst variants than hoechst 33342 solution 15. Use proper glove removal technique without touching gloves outer. Hoechst 33342 is a specific fluorescent dna stain that binds within the minor groove of doublestranded atrich regions. Hoechst 33342induced apoptosis in hl60 cellszhang et al 921. Invitrogen hoechst 33342, trihydrochloride, trihydrate.
Hoechst 33342 is a specific fluorescent dna stain that binds within the minor groove of doublestranded. The cells were suspended in hoechst 33342 solution 10. Hoechst 33342 are widely used for nuclear counterstaining, apoptosis and cell cycle studies. This safety data sheet complies with the requirements of regulation ec no. Fixation and permeabilization is not required for labeling cells, but physiologic conditions. This stain can be used on both live and fixed cells 1.
Hoechst 33342 is provided as a 10 mg solid c005 and a 10 mgml aqueous solution c006. It can also be used to detect the contents of a sample dna by plotting a standard emissiontocontent curve. It can be used to examine cellular dna in most fluorescent applications. Pureblu hoechst 33342 nuclear staining dye is a highly pure formulation of hoechst 33342 fluorescent dye figure 1 packaged in a userfriendly format. Catalog number pkca70740047 description hoechst dyes are cell membrane permeable and stain dna to emit intense blue fluorescence. Protocol labeling nuclear dna with hoechst 33342 1step 1, dilute the hoechst stock solution 1. Extraction of hoechst 33342 from the cytoplasmic leaflet of the plasma membrane by pglycoprotein article pdf available in european journal of biochemistry 2501.
Use of hoechst 33342 staining to detect apoptotic changes. Hoechst stains are part of a family of blue fluorescent dyes used to stain dna. Hoechst 33342, cas 23491523, is a high purity and quality chemical used as molecular probe and important as fluorescent stain for dna, chromosomes and nuclei. Cell cycle analysis using hoechst 33342 in unfixed cells date. Please observe safe laboratory practice and consult the safety datasheet. Quantitation of dna using hoechst 33258 an essential element of cellular and molecular biology is the ability to quantitate dna in large numbers of samples at a sensitivity that enables determination of small amounts of sample. Specific storage and handling information for each product is indicated on the product datasheet. Both hoechst 33342 and hoechst 33258 are watersoluble and stable in aqueous solutions. Loken 1980 simultaneous quantitation of hoechst 33342 and immunofluorescence on viable cells using a fluorescence activated cell sorter. Hoechst33342stainingforcellcycleanalysisoflivecells. Flow cytometric estimation of dna and rna content in.
Hoechst 33342 supplier cas 23491523 tocris bioscience. Hoechst 33342 is a popular cell membranepermeant nuclear counterstain. In vitro morphological assessment of apoptosis induced by. After incubation with hoechst 33342, the cells were counter stained with propidium iodide 2. Chromatin condensationdead cell apoptosis kit with.
Labeling nuclear dna with hoechst 33342 csh protocols. Simultaneous quantitation of hoechst 33342 and immunofluorescence on viable cells using a. The key difference between them is that the additional ethyl group of hoechst 33342 makes it more lipophilic, and thus more readily to cross intact cell membranes. Hoechst 33342 solution 20 mm thermo fisher scientific. Hoechst 33342 emits blue fluorescence upon binding to the minor groove of dsdna. Identification of the substancemixture and of the companyundertaking product name. Protocol cell assay 1 labeling nuclear dna with hoechst 33342 1step 1, dilute the hoechst stock solution 1. Hoechst 33342 nucleic acid stain is a popular cellpermeant nuclear counterstain that emits blue fluorescence when bound to dsdna. Hoechst 33342 is a bisbenzimidazole dye with high specificity for binding to doublestranded dna preferentially binds to at base pairs. Hoechst 33342 stainingforcellcycleanalysisoflivecells. Material safety data sheet hoechst 33342 3hcl bisbenzimide sc200908 hazard alert code key. Hoechst 33342 is a cellpermeable fluorescent compound that is able to stain the dna of eukaryotic and prokaryotic cells by binding with high affinity to the minor groove of atrich dna sequences. Hoechst dyes are cell membrane permeable and stain dna to emit intense blue fluorescence.
Our team of scientists has experience in all areas of research including life science, material science, chemical synthesis, chromatography, analytical and many others. For general guidelines, precautions, limitations on the use of our assay. V244 introduction apoptosis is a carefully regulated process of cell death that occurs as a normal part of. They bind to dna in the minor groove of polyat sequence rich areas.
It is found that the fluorescence emission of hoechst 33342 varies in low and high concentrations of dye, and the fluorescence of the cells shifts from blue to red with an increased dye intake, indicating a critical dyecell ratio for accurate hoechst 33342 staining petersen et al. Gfp transfection efficiency assay using hoechst 33342 and. The bd pharmingen hoechst 33342 solution is a reagent for the fluorescent staining of dna and nuclei in live or fixed cells. Identification of the substancemixture and of the companyundertaking product identifier product code h21492 product name hoechst 33342, trihydrochloride, trihydrate chemical name not applicable reach registration numberno registration number is given yet for this substance substances in this mixture.
Hoechst 33342 is a cellpermeable, benzimidazole dye that stains dna by binding to the minor groove of adenine and thyminerich sequences. Hoechst 33342 is used for specifically staining the nuclei of living or fixed cells. Identification of the substancemixture and of the companyundertaking product identifier product code h3570 product name hoechst 33342, trihydrochloride, trihydrate chemical name not applicable reach registration numberno registration number is given yet for this substance substances in this mixture. Product data sheet nb5117 pdf datasheet request cofa. Download product insert pdfdownload safety data sheet. Flow cytometric analysis of marine bacteria with hoechst 33342. This protocol describes the use of hoechst 33342 to label nuclear dna of cells grown in culture.
Bisbenzimide mixed with dna samples can then be used. Hoechst 33342 binds preferentially to adeninethymine at regions of dna. The purpose of this study was to determine whether m. Hoechst 33342 staining dye solution ab228551 abcam. Analyzing cell death by nuclear staining with hoechst 33342. Assignment of dna binding sites for 4,6diamidine2phenylindole and bisbenzimide hoechst 33258. In order to obtain this it is crucial to follow the protocol carefully. Hoechst 33342 can also be used to stain fixed cells by substituting hoechst 33342 for dapi1. Hoechst 33342 hydrochloride 875756971 cayman chemical. Hoechst 33342 is used for specifically staining the nuclei of living or fixed cells and tissues. This product may be used in fluorescence microscopy, microplate, cuvette, and flow cytometry applications. May be used for fluorescence microscopy or flow cytometry. It seemed likely that many of the problems arising from the multiple equilibria involving a0 might be eliminated by using separate stains for dna and rna. Approximate fl uorescence excitationemission maxima, when bound to dna.
The products may be used in fluorescence microscopy, microplate, cuvette, and flow. Thermo scientific pierce hoechst 33342 fluorescent stain is a highquality solution of hoechst dye for fixed and livecell fluorescent staining of dna and nuclei in cellular imaging techniques. In addition, the side population pattern is found to. Pdf extraction of hoechst 33342 from the cytoplasmic. Hoechst 33342 hydrochloride safety data sheet section 2. Catalog number pkca70740046 description hoechst dyes are cell membrane permeable and stain dna to emit intense blue fluorescence. Hoechst 33342, trihydrochloride, trihydrate 10 mgml. Hoechst 33342 handbook of fluorescent dyes and probes. Most apexbio products are stable under the recommended conditions. These bisbenzimides were originally developed by hoechst ag, which numbered all their compounds so that the dye hoechst 33342 is the 33,342nd compound made by the company.
Test principle hoechst 33342 is a premeable dna dye that binds preferentially to at basepairs. Bisbenzimide hoechst 33342 is an organic compound used as a fluorescent stain for dna in molecular biology applications. The assay depends on successful staining with solution 15 hoechst staining. This stain binds into the minor groove of dna and exhibits distinct fluorescence emission spectra that are dependent on dye. Hoechst 33342 induces apoptosis in hl60 cells and inhibits. It presents a list of imaginglabeling applications, biologicalmedical applications and industrial applications of hoechst 33342.
Step 2, aspirate the cell medium from cells grown on coverslips. This dye is often used to distinguish condensed pycnotic nuclei in apoptotic cells and for cell cycle studies in combination with brdu. Remove pbs and mount cells in antifademounting media. Hoechst dyeblue fluorescent stain specific for dna i. Hoechst stains 2 brdu quenching hoechst 33342 and 33258 are quenched by brdu. Hoechst 33342 20 mm solution in water 23491523 na 4. Reagent preparation briefly centrifuge small vials at low speed prior to opening. Product name hoechst 33342, trihydrochloride, trihydrate 10 mgml solution in water for research use only. The samples were maintained in the dark for 15 minutes. Products are sometimes shipped at a temperature that differs from the recommended storage temperature. Hoechst 33342 bisbenzimide h33342 trihydrochloride is supplied as a lyophilized powder in 25 mg units.
Using hoechst 33342 staining, we observed increased numbers of apoptotic monocytes. Hoechst 33342,nuclear counterstain, cell cycle genecopoeia. In some applications, hoechst 33258 is significantly less permeant than hoechst 33342. Safety data sheet in accordance with local and national regulations, the product is classified and labelled in accordance with directive 199945ec.
Flow cytometric analysis of marine bacteria with hoechst 33342 article pdf available in applied and environmental microbiology 593. View enhanced pdf access article on wiley online library html view download pdf for offline viewing. Hoechst 33342 hsc staining and stem cell purification protocol. Extreme high moderate low section 1 chemical product and company identification product name hoechst 33342 3hcl bisbenzimide statement of hazardous nature considered a hazardous substance according to osha 29 cfr 1910. Hoechst 33342 hydrochloride safety data sheet supersedes revision.
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