Hematoxylin

Manufactured by Roche

Sourced in United States, Germany

Hematoxylin is a dye commonly used in histology and microscopy as a staining agent. It is a natural dye derived from the wood of the Logwood tree (Haematoxylum campechianum). Hematoxylin stains cell nuclei and other structures, enabling the visualization of cellular and tissue components under a microscope.

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Lab products found in correlation

Bluing reagent, by Roche (9 mentions) Iview dab detection kit, by Roche (5 mentions) Discovery chromomap dab kit, by Roche (5 mentions) Cc1 buffer, by Roche (5 mentions) Benchmark xt staining module, by Roche (4 mentions) Ventana discovery ultra, by Roche (4 mentions) Discovery xt processor, by Roche (4 mentions) Ezprep buffer, by Roche (4 mentions) Dab detection kit, by Roche (4 mentions) Ultraview universal dab detection kit, by Roche (4 mentions) Cc1 standard benchmark xt pretreatment for antigen retrieval, by Roche (3 mentions) Permount, by Thermo Fisher Scientific (3 mentions) Ventana discovery xt, by Roche (3 mentions) Entellan, by Merck Group (2 mentions) Ultraview dab detection kit, by Roche (2 mentions) Synaptophysin, by Thermo Fisher Scientific (2 mentions) Chromogranin a, by Agilent Technologies (2 mentions) Biotinylated goat anti rat igg, by Vector Laboratories (2 mentions) Bond primary antibody diluent, by Leica (2 mentions) Dab map detection kit, by Roche (2 mentions)

52 protocols using hematoxylin

Immunohistochemical Analysis of Cadaveric Tissue

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Sections of paraformaldehyde fixed human tissue samples from fresh cadavers donated to the Department of Anatomy, Histology and Embryology of the Medical University of Innsbruck were stained with rabbit anti-ITGAV (ab179475, Abcam, UK), mouse anti-ITGA5 (MA5-15568, Thermo Scientific, Austria), mouse anti-Fibronectin (MS-1351, Thermo Scientific, Austria) or rabbit anti-PLIN1 ((D1D8) XP, cell signaling, USA) antibodies applying a Ventana Roche Discovery Immunostainer (Ventana, Germany) according to the DAB-MAP discovery research standard procedure. A biotinylated immunoglobulin cocktail of goat anti-mouse-IgG, goat anti-mouse-IgM, goat anti-rabbit-IgG and protein block (760–4205, Ventana) was applied for 30 min at room temperature. Hematoxylin (760–2021, Ventana) counterstained sections were manually dehydrated in downgraded alcohol series, cleared in xylene and permanently cover slipped with Entellan^® (Merck, Germany). Positive controls (placenta, liver) and negative control slides were added to each experiment to validate the IHC staining reactions. Digital images were acquired using AxioVision microscope software linked to an AxioCamHRc camera and an AxioPlan2 microscope (Zeiss, Germany). Because the dead bodies were immediately anonymized no certificate of non-objection was needed.

Morandi E.M., Verstappen R., Zwierzina M.E., Geley S., Pierer G, & Ploner C. (2016). ITGAV and ITGA5 diversely regulate proliferation and adipogenic differentiation of human adipose derived stem cells. Scientific Reports, 6, 28889.

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TUNEL Assay for Apoptosis Detection

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In situ detection of apoptotic cells was carried out using the TUNEL assay kit according to the manufacturer’s protocol (Millipore). Briefly, sections were deparaffinized and washed in PBS, followed by proteinase K pretreatment and endogenous peroxidase activity quenching. Labeling was performed by adding terminal deoxynucleotidyl transferase enzyme mix to the tissue sections, and the reaction was stopped by immersing slides in stop buffer followed by three PBS washes. After adding anti-digoxigenin conjugate to the slide, the color was developed using a diaminobenzidine peroxidase substrate kit (Vector Laboratories Inc.). Last, the slides were counterstained with hematoxylin (Ventana) and coverslipped. Images were captured with an Olympus DP70 camera attached to an Olympus BX51 microscope using a ×20 objective.

Chen S.H, & Yu X. (2019). Targeting dePARylation selectively suppresses DNA repair–defective and PARP inhibitor–resistant malignancies. Science Advances, 5(4), eaav4340.

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Immunohistochemical Analysis of Hepatocellular Carcinoma

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Tissue arrays were obtained from Superbiochips Laboratories (Seoul, Korea). The slide included specimens of HCC, metastatic HCC, and liver normal tissues obtained by biopsy or surgical resection from patients. Immunohistochemical staining was performed using a BenchMark XT automated slide stainer (Ventana Medical Systems, Inc., Tucson, AZ, USA) and OptiView DAB IHC detection Kit (Ventana). The slides were counterstained with hematoxylin (760-2021; Ventana Medical Systems, Inc., Tucson, AZ, USA) and Bluing reagent (760-2037; Ventana Medical Systems, Inc., Tucson, AZ, USA).

Yun U.J., Bae S.J., Song Y.R, & Kim Y.W. (2022). A Critical YAP in Malignancy of HCC Is Regulated by Evodiamine. International Journal of Molecular Sciences, 23(3), 1855.

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Automated ARPC1B Immunohistochemistry Protocol

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Formalin fixed tissues were dehydrated, embedded in paraffin and sectioned at 4 μm. A positive control was added on the right side of the slides. The slides were warmed up to 60°C for 1 hour, followed by fully automated processing. The ARPC1B immunostaining was calibrated on a Benchmark XT staining module (Ventana Medical Systems). Briefly, after sections were dewaxed and rehydrated, a CC1 Standard Benchmark XT pretreatment (Ventana Medical Systems) for antigen retrieval was selected. ARPC1B antibody (Novus Biologicals, USA, NBP1-90114) was diluted 1:100 and incubated 40 minutes at 37°C. Detection and counterstaining were performed with an ultraView detection kit (Ventana Medical Systems) and hematoxylin (Ventana Medical Systems), respectively. At the end of the automated run, slides were dehydrated by passage through increasing concentrations of ethanol. Sections were then cleared in xylene and mounted with Entellan followed by analysis by a pathologist.

Somech R., Lev A., Lee Y.N., Simon A.J., Barel O., Schiby G., Avivi C., Barshack I., Rhodes M., Yin J., Wang M., Yang Y., Rhodes J., Marcus N., Garty B.Z., Stein J., Amariglio N., Rechavi G., Wiest D.L, & Zhang Y. (2017). Disruption of thrombocyte and T lymphocyte development by a mutation in ARPC1B. Journal of immunology (Baltimore, Md. : 1950), 199(12), 4036-4045.

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Comprehensive Necropsy and Histological Analysis

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A complete necropsy was performed, and appropriate tissue samples were taken for histologic evaluation. All tissues were fixed in 10% neutral buffered formalin, processed conventionally, embedded in paraffin, cut at 5 microns, stained with Hematoxylin and eosin, and evaluated by light microscopy by at least one board-certified veterinary pathologist. Histochemical staining for periodic acid-Schiff (PAS), Gomori’s methenamine silver (GMS), Giemsa, and Gram’s stain were performed according to standard protocol. For immunohistochemical labeling, reagents were procured from Ventana Medical Systems, Tucson, AZ, USA. All stains were completed using the Ventana BenchMark Ultra automatic stainer (Ventana Medical Systems, Inc., Tucson, AZ, USA) with the UltraView DAB detection kit (Ventana Medical Systems, Cat # 760-500). Immunohistochemical labeling was performed on formalin-fixed, paraffin-embedded, 4 um thick sections of the thoracic mass as well as the myocardium. Slides were deparaffinized, and treated with Hier in CC1 at 95°C for 36 minutes, incubated with primary antibody, and counterstained with Hematoxylin (Ventana catalog number 760–2021)/Bluing (Ventana catalog number 760–2037) for 4 minutes each. The specific primary antibodies and their respective incubation temperatures are listed in Table 1.

Morosco D.T., Cline C.R., Owston M.A., Kumar S, & Dick EJ J.r. (2017). Spontaneous mediastinal myeloid sarcoma in a common marmoset (Callithrix jacchus) and review of the veterinary literature. Journal of medical primatology, 46(2), 42-47.

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Immunohistochemical Analysis of Bone Tumors

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The fixed bone tumors were decalcified for 7 d before embedding in paraffin. The embedded tibias and lungs were sectioned and stained with H&E, or IHC for CD99 (ARUP Laboratories, Salt Lake City, UT). The expression of CD99 was detected using clone O-13 anti-mouse antibody (1:200 dilution) for 2 h at 37°C. Biotinylated secondary antibody (1:100) was applied for 32 min at 37°C. Signals were detected with a streptavidin-HRP system, using 3-3′ diaminobenzidine as the chromogen (Research DAB Detection Kit; Ventana Medical Systems, Tucson, AZ). The slides were counterstained with hematoxylin (Ventana Medical Systems) for 8 min, gently washed in deionized H₂O/Dawn detergent mixture, placed in iodine for 30 s, then dipped in sodium thiosulfate, dehydrated in graded alcohols (70%, 95% ×2, and 100% ×2), and cleared in xylene; coverslips were then mounted (Sheryl Tripp at ARUP and Y.-C.L. at the Huntsman Cancer Institute). The tibia and lung sections were evaluated by an independent clinical pathologist (A.G.) in a blind study.

Chaturvedi A., Hoffman L.M., Jensen C.C., Lin Y.C., Grossmann A.H., Randall R.L., Lessnick S.L., Welm A.L, & Beckerle M.C. (2014). Molecular dissection of the mechanism by which EWS/FLI expression compromises actin cytoskeletal integrity and cell adhesion in Ewing sarcoma. Molecular Biology of the Cell, 25(18), 2695-2709.

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Immunohistochemical Staining of FFPE Samples

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FFPE blocks were sectioned at a thickness of 4μm and a positive control was added on the right side of the slides. All immunostainings were fully calibrated on a Benchmark XT staining module (Ventana Medical Systems Inc., USA). Briefly, after sections were dewaxed and rehydrated, a CC1 Standard Benchmark XT pretreatment for antigen retrieval (Ventana Medical Systems) was selected for all immunostainings: Chromogranin A (1:500, Dako, Denmark), Synaptophysin, (1:200, Life Technologies, Invitrogen, USA), GAST, CGC and STT. Detection was performed with iView DAB Detection Kit (Ventana Medical Systems Inc., USA) and counterstained with hematoxylin (Ventana Medical Systems Inc., USA). After the run on the automated stainer was completed, slides were dehydrated in ethanol solutions (70%, 96%, and 100%) for one min each. Sections were then cleared in xylene for 2 min, mounted with Entellan and cover slips were added.

Oz-Levi D., Olender T., Bar-Joseph I., Zhu Y., Marek-Yagel D., Barozzi I., Osterwalder M., Alkelai A., Ruzzo E.K., Han Y., Vos E.S., Reznik-Wolf H., Hartman C., Shamir R., Weiss B., Shapiro R., Pode-Shakked B., Tatarskyy P., Milgrom R., Schvimer M., Barshack I., Imai D.M., Coleman-Derr D., Dickel D.E., Nord A.S., Afzal V., van Bueren K.L., Barnes R.M., Black B.L., Mayhew C.N., Kuhar M.F., Pitstick A., Tekman M., Stanescu H.C., Wells J.M., Kleta R., de Laat W., Goldstein D.B., Pras E., Visel A., Lancet D., Anikster Y, & Pennacchio L.A. (2019). Noncoding Deletions Expose a Novel Gene Critical for Intestinal Function. Nature, 571(7763), 107-111.

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Immunohistochemical Analysis of 3D Cultures

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The 3D cultures were fixed in 10% neutral buffered formalin, paraffin embedded, sectioned using a microtome, and stained with H&E, according to commonly used procedures. The IHC stain was performed on a Ventana Discovery Ultra (Ventana Medical Systems, Roche Diagnostics, Indianapolis, IN, USA) automated IHC stainer. The slides were loaded onto the machine, and deparaffination, rehydration, endogenous peroxydase activity inhibition, and antigen retrieval were performed. Following primary antibody incubation, DISCOVERY anti-Rabbit HQ and DISCOVERY anti-HQ-HRP were incubated. The stains were then visualized with the DISCOVERY ChromoMap DAB Kit, counterstained with hematoxylin (Ventana). Primary antibody used: Ki-67: Clone# 30-9, rabbit monoclonal antibody; P53: Clone# DO-7, mouse monoclonal antibody (Ventana). Images were taken with a VENTANA iScan HT slide scanner and analyzed with a Ventana Image Viewer.

Chen M., Liang S., Shahid A., Andresen B.T, & Huang Y. (2020). The β-Blocker Carvedilol Prevented Ultraviolet-Mediated Damage of Murine Epidermal Cells and 3D Human Reconstructed Skin. International Journal of Molecular Sciences, 21(3), 798.

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Immunohistochemical Localization of Reg3β

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IHC detection of Reg3β was performed at the Molecular Cytology Core Facility of Memorial Sloan Kettering Cancer Center using a Discovery XT processor (Ventana Medical Systems). Formalin-fixed tissue sections were deparaffinized with EZPrep buffer (Ventana Medical Systems), antigen retrieval was performed with CC1 buffer (Ventana Medical Systems) and sections were blocked for 30 minutes with Background Buster solution (Innovex). Slides were incubated with anti-Reg3β antibodies (R&D Systems; cat# MAB5110; 1ug/ml) or isotype (5ug/ml) for six hours, followed by a 60 minute incubation with biotinylated goat anti-rat IgG (Vector labs, cat# PK-4004) at 1:200 dilution. The detection was performed with DAB detection kit (Ventana Medical Systems) according to the manufacturer's instructions. Slides were counterstained with hematoxylin (Ventana Medical Systems) and coverslipped with Permount (Fisher Scientific). Refer to Supplementary Table 1 for full description of antibodies used.

Lindemans C.A., Calafiore M., Mertelsmann A.M., O'Connor M.H., Dudakov J.A., Jenq R.R., Velardi E., Young L.F., Smith O.M., Lawrence G., Ivanov J.A., Fu Y.Y., Takashima S., Hua G., Martin M.L., O'Rourke K.P., Lo Y.H., Mokry M., Romera-Hernandez M., Cupedo T., Dow L., Nieuwenhuis E.E., Shroyer N.F., Liu C., Kolesnick R., van den Brink M.R, & Hanash A.M. (2015). Interleukin-22 Promotes Intestinal Stem Cell-Mediated Epithelial Regeneration. Nature, 528(7583), 560-564.

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Lung Tissue Histology Workflow

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Lung tissues were harvested and immediately fixed in 10% neutral buffered formalin. Dehydration, clear, and paraffinization were performed on a Tissue‐Tek VIP Vacuum Infiltration Processor (SAKURA). The samples were embedded in paraffin using a Tissue‐Tek TEC Tissue Embedding Station (SAKURA). Samples were then sectioned at 5 μm and put on positively charged glass slides. The slides were deparaffinized, rehydrated, and stained with Modified Mayer's hematoxylin and Eosin Y (H&E) Stain (America MasterTech Scientific) on an H&E Auto Stainer (Prisma Plus Auto Stainer, SAKURA) according to standard laboratory procedures. The stains were visualized with DISCOVERY ChromoMap DAB Kit, counterstained with hematoxylin (Ventana) and cover‐slipped. H&E stained slides were digitalized and documented by iScan HT (Roche) scanner, and then representative pictures (10× magnified) were taken from the NDP.view2 viewing software.

Williams L., Li L., Yazaki P.J., Wong P., Hong T., Poku E.K., Hui S., Ghimire H., Shively J.E, & Kujawski M. (2024). Comparison of IL‐2‐antibody to IL‐2‐Fc with or without stereotactic radiation therapy in CEA immunocompetent mice with CEA positive tumors. Cancer Medicine, 13(3), e6909.

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