Hoechst dna dye

Manufactured by Thermo Fisher Scientific

Hoechst DNA dye is a fluorescent stain used to detect and quantify DNA. It binds to the minor groove of double-stranded DNA, emitting a blue fluorescence upon excitation. The dye is commonly used in various applications, such as flow cytometry and fluorescence microscopy, to visualize and analyze DNA content in cells.

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Lab products found in correlation

Alexa fluor 647 conjugated goat anti rabbit secondary antibody, by Thermo Fisher Scientific (1 mentions) Citrate buffer, by Merck Group (1 mentions) Prolong mounting medium, by Thermo Fisher Scientific (1 mentions) Triton x, by Merck Group (1 mentions) Mouse anti aβ, by Cell Signaling Technology (1 mentions) Rat anti mouse cd8a, by Thermo Fisher Scientific (1 mentions) Alexa fluor secondary antibody, by Thermo Fisher Scientific (1 mentions) Thioflavin s, by Merck Group (1 mentions)

Close spelling variants of this product

DNA dye Hoechst 33342 Hoechst dye Hoechst

2 protocols using hoechst dna dye

Quantifying SARS-CoV-2 Infection by Immunofluorescence

Cited 1 time

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Fixed cells were washed three times with Dulbecco-modified PBS containing 0.2% BSA (DPBS/BSA), permeabilized with 0.1% Triton X-100 in DPBS/BSA and processed for immunodetection of viral N protein, automated fluorescence imaging, and image analysis. Briefly, viral NP was detected with an in-house-developed rabbit polyclonal antibody^{22 (link)} counterstained with Alexa Fluor 647-conjugated goat anti-rabbit secondary antibody (ThermoFisher Scientific, catalogue number A32733); nuclear staining was done using Hoechst DNA dye (ThermoFisher Scientific, catalogue number H3570). Automated fluorescence imaging was done using a Molecular Devices Image-Xpress Nano high-content epifluorescence microscope equipped with a 10× objective and a 4.7-megapixel CMOS camera (pixel size, 0.332 μm). Image analysis was performed with CellProfiler-4 software (www.cellprofiler.org). Automated detection of nuclei was performed using the Otsu algorithm inbuilt in the software. To automatically identify infected cells, an area surrounding each nucleus (5-pixel expansion of the nuclear area) was used to estimate the fluorescence intensity of the viral NP immunolabeled protein, using an intensity threshold such that <0.01% of ‘positive cells’ were detected in noninfected wells.

Kant R., Kareinen L., Ojha R., Strandin T., Saber S.H., Lesnikova A., Kuivanen S., Sirnonen T., Joensuu M., Vapalahti O., Kirchhausen T., Kipar A, & Balistreri G. (2023). Complete Protection from SARS-CoV-2 Lung Infection in Mice Through Combined Intranasal Delivery of PIKfyve Kinase and TMPRSS2 Protease Inhibitors. bioRxiv.

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Immunohistochemical Analysis of Brain Tissues

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Paraffin-embedded brain tissues were sectioned at 5-μm thickness. Deparaffinization was achieved by washing slides through a series of xylenes and decreasing concentrations of ethanol. Tissue sections were then subjected to antigen retrieval using citrate buffer, pH 6.0 (Sigma-Aldrich) at 95 °C for 30 min. Following rinsing with PBS, sections were incubated in blocking buffer containing PBS with 10% normal donkey serum and 0.03% Triton-X (Sigma-Aldrich) for 2 h at room temperature. Slides were then incubated with primary antibody in blocking buffer overnight at 4°C. The following day, slides were rinsed with PBS then incubated in appropriate Alexa Fluor secondary antibodies (Thermo Fisher Scientific). Sections were then rinsed and stained with Hoechst DNA dye (Thermo Fisher Scientific) before being coverslipped with ProLong mounting medium (Invitrogen). Primary antibodies included rat anti-human CD3 (Abcam), rabbit anti-human CD8α (Cell Signaling), mouse anti-Aβ (Cell Signaling), chicken anti-human MAP2 (Abcam), mouse anti-human granzyme-A (Abcam), rat anti-mouse CD8a (eBioscience) and rabbit anti-mouse NEFH (Abcam). For mouse Aβ plaque staining, ThioflavinS (1 mg ml⁻¹, 1:625, Sigma) was added to the secondary antibody solution.

Gate D., Saligrama N., Leventhal O., Yang A.C., Unger M.S., Middeldorp J., Chen K., Lehallier B., Channappa D., De Los Santos M.B., McBride A., Pluvinage J., Elahi F., Tam G.K., Kim Y., Greicius M., Wagner A.D., Aigner L., Galasko D.R., Davis M.M, & Wyss-Coray T. (2020). Clonally expanded CD8 T cells patrol the cerebrospinal fluid in Alzheimer’s disease. Nature, 577(7790), 399-404.

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