Hoechst fluorescent stain

Manufactured by Thermo Fisher Scientific

Hoechst fluorescent stain is a nucleic acid stain used to detect and quantify DNA in various applications. It binds to the minor groove of DNA, emitting a blue fluorescent signal upon excitation. The stain can be used with various microscopy techniques and flow cytometry for the analysis of cellular DNA content.

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Lab products found in correlation

Fv10i scanning confocal microscope, by Olympus (1 mentions) Alexa 594 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Primary antibody against p65, by Cell Signaling Technology (1 mentions) Adipor1, by Merck Group (1 mentions) Lsr 2 flow cytometer, by BD (1 mentions)

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Hoechst (1591 citations) Hoechst stain (133 citations) Hoechst 33342 fluorescent stain (15 citations)

2 protocols using hoechst fluorescent stain

Immunofluorescence Analysis of BV2 Cells

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BV2 cells were grown on poly-D-lysine-coated glass coverslips. After treatment, cells were fixed with 3.2% paraformaldehyde, then permeabilized in PBS pH 7.4, containing 0.3% Tween and 3% BSA. Cells were then incubated overnight with the appropriate primary antibody directed against CD11b (Abcam), iNOS (Merck Millipore), AdipoR1 or AdipoR2 (Sigma Aldrich) in the same buffer. At the end of the incubation time, cells were washed three times with PBS and incubated with either Alexa488-conjugated or Alexa594-conjugated secondary antibodies (Invitrogen) and Hoechst fluorescent stain (Invitrogen) for 1 h at room temperature. For NF-κB p65 subunit nuclear translocation study, cells were pre-treated with 10 μg/ml gApN or saline for 1 h before addition of 0.5 μg/ml LPS for 3 additional hours. Cells were then fixed and permeabilized as previously. Primary antibody against p65 was from Cell Signaling. Images were captured with an FV10i scanning confocal microscope (Olympus, France) and analyzed using Image J software (“National Institutes of Health” Image J Software™).

Nicolas S., Cazareth J., Zarif H., Guyon A., Heurteaux C., Chabry J, & Petit-Paitel A. (2017). Globular Adiponectin Limits Microglia Pro-Inflammatory Phenotype through an AdipoR1/NF-κB Signaling Pathway. Frontiers in Cellular Neuroscience, 11, 352.

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Live cell gGlu-HMRG imaging of SCC

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The protocol adopted for this study was described previously by Urano et al. [12 ]. Flow cytometry was performed with the BD LSR-II Flow Cytometer for cell evaluation and subsequent cell measurements were analysed with FlowJo software (FlowJo, LLC, Ashland, OR, USA). Two human SCC cell lines, OSC19 (oral cancer; GGT overexpressing) and FaDu (hypopharyngeal cancer; GGT low expressing), were used. After incubation with gGlu-HMRG, the cells were nuclear stained with Hoechst fluorescent stain (Invitrogen). Images were obtained every 5 min for 1 h after application using the Leica microsystems LAS AF6000 modular systems at 20× magnification. Quantification of the signal over time was assessed using the in-house developed software program, called ‘Stacks’, which operates using the Microsoft Windows operating system. Each measurement was corrected to make translation between separate images possible after which a mask was created based on the last image (t = ’60). Within the mask the average intensity was measured for all images and the standard deviation was calculated as a percentage of the intensity.

Slooter M.D., Handgraaf H.J., Boonstra M.C., van der Velden L.A., Bhairosingh S.S., Que I., de Haan L.M., Keereweer S., van Driel P.B., Chan A., Kobayashi H., Vahrmeijer A.L, & Löwik C.W. (2018). Detecting tumour-positive resection margins after oral cancer surgery by spraying a fluorescent tracer activated by gamma-glutamyltranspeptidase. Oral oncology, 78, 1-7.

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