Hoechst

Manufactured by Roche

Sourced in United States

The Hoechst lab equipment is a versatile instrument used for various applications in research and clinical settings. It provides a core function of fluorescence detection and analysis. The Hoechst performs this task through its advanced optical components and integration with compatible software systems.

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Lab products found in correlation

Nitroblue tetrazolium, by Merck Group (2 mentions) Na succinate, by Merck Group (2 mentions) Dpx compound, by Merck Group (2 mentions) Alexa fluor 488 conjugate, by Thermo Fisher Scientific (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Roche (2 mentions) Vs110, by Olympus (1 mentions) Igg mouse, by Santa Cruz Biotechnology (1 mentions) Igg rabbit, by Santa Cruz Biotechnology (1 mentions) Cy3 or cy5, by Jackson ImmunoResearch (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) E cadherin, by Thermo Fisher Scientific (1 mentions) Ssea 1, by Developmental Studies Hybridoma Bank (1 mentions) β catenin, by Thermo Fisher Scientific (1 mentions) Axin2, by Santa Cruz Biotechnology (1 mentions) Ficin, by Thermo Fisher Scientific (1 mentions) Cleaved parp, by Cell Signaling Technology (1 mentions) β catenin, by Cell Signaling Technology (1 mentions) Vectashield, by Vector Laboratories (1 mentions) Wheat germ agglutinin (wga), by Thermo Fisher Scientific (1 mentions) Poly lysine objective slides, by Avantor (1 mentions)

Close spelling variants of this product

Hoechst 33342 Hoechst 33258

6 protocols using hoechst

Muscle Fiber Composition and Morphology Analysis

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TA and TB muscles were dissected out and snap-frozen in liquid nitrogen. For the muscle fiber composition (SDH staining), 10 μm-thickness serial coronal cryosections from the mid-belly region of the TA muscle were air-dried and then incubated at 37°C for 30 min in phosphate buffer (0.2 M, pH 7.6) containing 13.5 mg/mL Na-succinate (Sigma-Aldrich) and 0.5 mg/mL nitro blue tetrazolium (Sigma-Aldrich, 0.29 mg/mL buffer solution). After staining, sections were fixed with 4% paraformaldehyde, dehydrated in 15% alcohol for 5 min, and finally mounted with DPX compound (Sigma-Aldrich).
For the muscle fiber cross-sectional area, 10-μm thickness serial coronal cryosections from the mid-belly region of the TA muscle were air-dried, fixed in 4% paraformaldehyde solution for 5 min, and stained with wheat germ agglutinin, Alexa Fluor 488 conjugate (1:500; W11261, Thermo Fisher, Pittsburgh, PA, USA) and Hoechst (1:1,000; Roche).
Images were acquired with an Olympus virtual slide system VS110 (Olympus, Center Valley, USA) at 20× magnification and analyzed through Fiji (ImageJ) on 3–5 serial sections per animal. For the SDH staining, a systematic random sampling procedure was applied as described above. For the muscle fiber cross-sectional area and centralized nuclei, the entire TA or TB muscle section was analyzed with the MuscleJ plug-in of Fiji software, as previously described.¹⁴¹ (link)

Trolese M.C., Scarpa C., Melfi V., Fabbrizio P., Sironi F., Rossi M., Bendotti C, & Nardo G. (2022). Boosting the peripheral immune response in the skeletal muscles improved motor function in ALS transgenic mice. Molecular Therapy, 30(8), 2760-2784.

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Antibody Characterization for Cell Biology

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The InlP polyclonal antibody was obtained after injection of an immunogenic peptide (amino acids 365 to 379, LDVSYNHNYATGGVC) in rabbits and subsequent affinity purification. The InlC polyclonal antibody is described in^{27 (link)}. The other primary antibodies are polyclonal antibodies against RBM5 (Sigma, HPA017335 for IF and Bethyl, A302-228A for IP), PML (Abcam, sc-966), anticoilin (Proteintech, 10967-1-AP), anti-nucleolin (Santa cruz, sc-13057) and monoclonal antibodies against tubulin (hybridoma E7), SC35 (AbCam, ab11826), FLAG-M2 (Sigma-Aldrich, F1804), Myc (9E10, Santa Cruz Biotechnology, sc-40), HA (6E2, Cell Signaling technology #2367), V5 (R960-2, Invitrogen). The immunoprecipitation control antibodies are IgG mouse (Santa-Cruz, sc-2025) and IgG rabbit (Santa-Cruz sc-2027). The secondary antibodies are coupled to Alexa-488 (Life technologies) or Cy3 or Cy5 (Jackson ImmunoResearch). DAPI and Hoechst are from Roche Applied Sciences and Thermo Fisher Scientific, respectively. Lipofectamine LTX Max is from Invitrogen.

Pourpre R., Lakisic G., Desgranges E., Cossart P., Pagliuso A, & Bierne H. (2022). A bacterial virulence factor interacts with the splicing factor RBM5 and stimulates formation of nuclear RBM5 granules. Scientific Reports, 12, 21961.

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Immunohistochemical Analysis of Embryonic Markers

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Embryos fixed in 4% PFA were embedded in OCT and cryosectioned at a thickness of 10 µm. Slides were blocked 1 h in 10% calf serum + 0.1% Triton X-100 in PBS and stained overnight at 4°C in the blocking buffer followed by three 15-min washes in PBS. Primary antibodies used included β-catenin (rabbit, 1:25; #9582s; Cell Signaling Technology), Axin2 (goat, Conductin M-20, 1:100; #sc-8570; Santa Cruz Biotechnology, Inc.), GFP (chicken, Aves GFP-1020, 1:200), SSEA1 (mouse IgM, 1:200; MC-480; Developmental Studies Hybridoma Bank), Vasa (rabbit, 1:400; #ab13840-100; Abcam), cleaved PARP (rabbit, 1:50; #9544s; Cell Signaling Technology), E-cadherin (rat, 1:200; #13–1900; Invitrogen), Wnt5a (goat, 1:20; #AF645; R&D Systems), and Stella (rabbit, 1:100; #ab19878; Abcam). Histological staining for β-catenin was preceded by 10 min additional fixation in 4% PFA and 3 min treatment in undiluted Ficin (Invitrogen) at room temperature followed by a quick PBS wash. This treatment was not necessary for β-catenin staining on cultured cells. EdU was labeled per kit protocol (#C10338 or C10340; Thermo Fisher Scientific). Secondary antibodies were purchased from Invitrogen and incubated for 1 h in blocking buffer at room temperature at 1:200. Nuclei were labeled with DAPI or Hoechst (1:1,000; Roche or Sigma-Aldrich). Sections were mounted in Vectashield (Vector Laboratories).

Cantú A.V., Altshuler-Keylin S, & Laird D.J. (2016). Discrete somatic niches coordinate proliferation and migration of primordial germ cells via Wnt signaling. The Journal of Cell Biology, 214(2), 215-229.

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Muscle Composition and Fiber Analysis

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TA muscles were dissected out and snap‐frozen in liquid nitrogen. Serial transverse cryosections (10 μm) were air‐dried, fixed in 4% paraformaldehyde solution for 5′, and stained with Wheat Germ Agglutinin, Alexa Fluor™ 488 Conjugate (1:500; Thermo Fisher) and Hoechst (1:1000; Roche).
For the muscle composition, serial transverse cryosections (10 μm) were air‐dried and then incubated at 37°C for 30′ in phosphate buffer (0.2 M, pH 7.6) containing 13.5 mg/mL Na‐succinate (Sigma‐Aldrich, St. Louis, MO, USA) and 0.5 mg/mL of nitro blue tetrazolium (Sigma‐Aldrich, 0.29 mg/mL of buffer solution). After staining, sections were fixed with 4% paraformaldehyde, dehydrated in 15% alcohol for 5′ and finally mounted with DPX compound (Sigma Aldrich).
Images were acquired with an Olympus virtual slide system VS110 (Olympus, Center Valley, USA) at 20×‐magnification and analyzed through Fiji (Image J, U.S. National Institute of Health, Bethesda, Maryland, USA) on three serial sections per animal.

Fabbrizio P., Apolloni S., Bianchi A., Salvatori I., Valle C., Lanzuolo C., Bendotti C., Nardo G, & Volonté C. (2019). P2X7 activation enhances skeletal muscle metabolism and regeneration in SOD1G93A mouse model of amyotrophic lateral sclerosis. Brain Pathology, 30(2), 272-282.

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Lung Tissue Analysis in Anesthetized Mice

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Mice were deeply anesthetized with Medetomidine, 2 mg kg⁻¹, and Ketamin, 150 mg kg^−1, and transcardially perfused with 50 mL of 0.1 M PBS. The lungs were harvested and snap-frozen in liquid nitrogen; 20 μm longitudinal serial lung cryosections were collected on polylysine objective slides (VWR International). Sections were fixed in cold acetone for 10 minutes, then blocked in PBS with 10% normal goat serum and 0.3% Triton X-100, for 1 hour at room temperature. Sections were incubated overnight with primary antibodies: anti-CD11b, rat (1:200; BioRad) or anti-Ki67, rabbit (1:200; Abcam) at 4°C, then stained with the secondary antibody Alexa 488 anti-rat or anti-rabbit. The nuclei were counterstained whit Hoechst (1:1000; Roche). Images were acquired using a sequential scanning mode with an A1 Nikon confocal running NIS Elements at 20X or 40X magnification. Image analyses were done as detailed in Supplemental Methods.

Russo M., Panini N., Fabbrizio P., Formenti L., Becchetti R., Matteo C., Meroni M., Nastasi C., Cappelleri A., Frapolli R., Nardo G., Scanziani E., Ponzetta A., Bani M.R., Ghilardi C, & Giavazzi R. (2023). Chemotherapy-induced neutropenia elicits metastasis formation in mice by promoting proliferation of disseminated tumor cells. Oncoimmunology, 12(1), 2239035.

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EdU Incorporation and Hoechst Staining

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Cells were incubated with 50 μm EdU (Ribo-Bio, Guangzhou, China) for 12 h and then fixed with 4% paraformaldehyde for 30 min at 25 °C. Next, the cells were washed in PBS (2 × 5 min, room temperature [RT]) and then permeabilized using PBS containing 0.3% Triton X-100 for 10 min. After extensive washing in PBS, the cells were incubated in Apollo staining solution (RiboBio) for 20 min, washed with NaCl/Pi (3 × 10 min, RT), and then incubated in Hoechst (1:2500; Roche Diagnostics, Mannheim, Germany) for 10 min at RT.

Zhang H., Deng T., Ge S., Liu Y., Bai M., Zhu K., Fan Q., Li J., Ning T., Tian F., Li H., Sun W., Ying G, & Ba Y. (2018). RETRACTED ARTICLE: Exosome circRNA secreted from adipocytes promotes the growth of hepatocellular carcinoma by targeting deubiquitination-related USP7. Oncogene, 38(15), 2844-2859.

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