Hematoxylin gill no 3

Manufactured by Merck Group

Sourced in United States

Hematoxylin (Gill No. 3) is a laboratory stain used in histology and cytology. It is a natural dye extracted from the logwood tree and is commonly used to stain nucleic acids in biological samples, providing a deep blue-purple color.

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Lab products found in correlation

Hrp conjugated goat anti rabbit igg, by Santa Cruz Biotechnology (2 mentions) Eclipse ts100, by Nikon (2 mentions) Cellsens dimensions 2, by Olympus (1 mentions) Roti histol, by Merck Group (1 mentions) Eosin y, by Merck Group (1 mentions) Protein block solution, by Agilent Technologies (1 mentions) Antigen retrieval solution, by Agilent Technologies (1 mentions) Xylene, by Thermo Fisher Scientific (1 mentions) Phosphate buffer saline (pbs), by Thermo Fisher Scientific (1 mentions) Hydrogen peroxide, by Thermo Fisher Scientific (1 mentions) Immunocruz goat abc staining system, by Santa Cruz Biotechnology (1 mentions) 3 3 diaminobenzidine (dab), by Vector Laboratories (1 mentions) Anti luciferase antibody, by Abcam (1 mentions) Bloxall blocking solution, by Vector Laboratories (1 mentions) R buffer a, by Electron Microscopy Sciences (1 mentions) Ix73 inverted microscope, by Olympus (1 mentions) Streptavidin horseradish peroxidase, by Agilent Technologies (1 mentions) Anti rabbit secondary antibody, by Agilent Technologies (1 mentions) Reveal decloaker, by Biocare Medical (1 mentions)

7 protocols using hematoxylin gill no 3

Hematoxylin and Eosin Staining Protocol

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Specimens were stained for 3 min in hematoxylin (Gill No. 3, Sigma-Aldrich, Munich, Germany), washed 2 times with H₂O followed by differentiating in acidic alcohol (1% HCl in ethanol) for 3 s and bluing under running tap water for 10 min. Counterstaining was conducted with eosin (Eosin Y, Sigma-Aldrich, Munich, Germany) for 10 s. Dehydration was performed in increasing alcohol series and Roti-Histol^®. Finally, sections were mounted with Entellan^®. One area per animal was evaluated based on the region of trauma. Six animals were used for each group based on the power analysis conducted for the animal experiment application (license number: 1183). Pictures were taken with the UC30 color camera at X10 and X40 magnification. All stainings were acquired with an Olympus IX81 and analyzed with the Olympus software cellSens Dimensions 2.3 (Build 18987).

Gihring A., Gärtner F., Liu C., Hoenicka M., Wabitsch M., Knippschild U, & Xu P. (2020). Influence of Obesity on the Organization of the Extracellular Matrix and Satellite Cell Functions After Combined Muscle and Thorax Trauma in C57BL/6J Mice. Frontiers in Physiology, 11, 849.

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Immunohistochemical Analysis of Apoptosis Markers

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Rat testis tissue was fixed in Bouin’s solution and embedded in paraffin, and cut into sections with a thickness of 5 μm that were collected on glass slides. Sections were deparaffinized in xylene, rehydrated through graded series of ethanol, and rinsed with water [37 (link)]. Endogenous peroxidase activity was blocked by incubation in 0.3% hydrogen peroxide in PBS for 30 min at room temperature. Slides were blocked for 1 h in PBS supplemented with 10% normal goat serum. Bcl-2, Bax, and cleaved caspase-3 expression was detected by overnight incubation at 4°C with antibodies against Bax (1:100), Bcl-2 (1:50), or cleaved caspase-3 (1:100). After washing, sections were incubated for 1 h with HRP-conjugated goat anti-rabbit IgG (1:2000; Santa Cruz Biotechnology, USA) in 10% goat serum, then counterstained for 10 s with hematoxylin (Gill no. 3; Sigma, USA) and examined with a light microscope (Nikon Eclipse TS100, Japan).

Zhang K., Ge Z., Fu L., An Q., Zhou F., Guo Y., Wang X., Lu W., Liang X., Wang S., Shang X, & Gu Y. (2018). Qilin pills alleviate oligoasthenospermia by inhibiting Bax-caspase-9 apoptosis pathway in the testes of model rats. Oncotarget, 9(31), 21770-21782.

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Immunohistochemical Staining of CITED2 in Formalin-Fixed Tissues

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Formalin-fixed and paraffin-embedded tissue sections were deparaffinized in xylene (Fisher Scientific) and rehydrated through a graded series of ethanol (Pharmco-AAPER). Sections were immersed in antigen retrieval solution (Dako) and heated in a steamer for 20 minutes. Cooled sections were washed with phosphate buffer saline (PBS, Gibco) and endogenous peroxidase activity was quenched by immersing sections in 3% hydrogen peroxide (Fischer Scientific) for 12 minutes and washed with PBS. Sections were blocked by incubation with protein block solution (Dako) for 30 minutes at room temperature and incubated at 4°C for 18 hours with goat anti-CITED2 (1:500; Everest Biotech). Sections were then sequentially incubated for 15 minutes at room temperature with streptavidin-biotin complex, Tyramide amplification reagent and streptavidin-horse radish peroxidase (HRP) from the DACO CSA Kit (Vector Laboratories). To visualize proteins, the chromogen 3, 3-diaminobenzamindine (DAB; Open Biosystems) was added for two minutes at room temperature. Sections were subsequently washed in water and counterstained with hematoxylin Gill No. 3 (Sigma-Aldrich).

Jayaraman S., Doucet M., Lau W.M, & Kominsky S.L. (2016). CITED2 Modulates Breast Cancer Metastatic Ability Through Effects on IKKα. Molecular cancer research : MCR, 14(8), 730-739.

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Immunohistochemical Analysis of TSSK2 in Rat Testis

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Rat testis tissue was fixed by soaking in Bouin fixing fluid at room temperature for 48 h, and they then were dehydrated, embedded in paraffin, and cut into 5-μm-thick sections that were collected on glass slides. The sections were deparaffinized in xylene, rehydrated through a graded series of ethanol, and rinsed with water. Endogenous peroxidase activity was blocked by incubation in 0.3% hydrogen peroxide in PBS for 30 min at room temperature. The slides were blocked for 1 h in PBS supplemented with 10% normal goat serum. TSSK2 expression was detected after overnight incubation at 4 °C with antibodies against TSSK2 (1:200, Abcam, USA). After washing, the sections were incubated for 1 h with HRP-conjugated goat anti-rabbit IgG (1:2000; Santa Cruz Biotechnology, USA) in 10% goat serum, counterstained for 10s with hematoxylin (Gill no. 3; Sigma, USA). Imaging analyses was conducted using a confocal microscope (Nikon Eclipse TS100, Japan).

Zhang K., Fu L., An Q., Hu W., Liu J., Tang X., Ding Y., Lu W., Liang X., Shang X, & Gu Y. (2020). Effects of Qilin pills on spermatogenesis, reproductive hormones, oxidative stress, and the TSSK2 gene in a rat model of oligoasthenospermia. BMC Complementary Medicine and Therapies, 20, 42.

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Immunolocalization of Seipin in HepG2 Cells

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HepG2 cells were fixed on slides with 4% paraformaldehyde for 30min at RT. Fixed HepG2 cells, were blocked for one hour in 1x PBS containing 1.5% normal goat serum.
Samples were incubated with anti-Seipin primary antibody (Santa Cruz Biotechnology, clone L-16; 1:50) overnight at 4°C before using the ImmunoCruz goat ABC staining system (Santa Cruz Biotechnology). Nuclei were stained with Hematoxylin Gill no°3 (Sigma-Aldrich).

Lounis M.A., Lalonde S., Rial S.A., Bergeron K.F., Ralston J.C., Mutch D.M, & Mounier C. (2017). Hepatic BSCL2 (Seipin) Deficiency Disrupts Lipid Droplet Homeostasis and Increases Lipid Metabolism via SCD1 Activity. Lipids, 52(2).

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Immunohistochemistry of Luciferase in FFPE Tissues

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IHC staining on formalin-fixed, paraffin-embedded tissue sections was performed as before (18 (link)). Briefly, slides were deparaffinized and rehydrated, epitope retrieval was performed in a Retriever 2100 (Aptum Biologics) with R-Buffer A (Electron Microscopy Sciences), and endogenous peroxidases/phosphatases were quenched with BLOXALL blocking solution (Vector Laboratories). Tissues were blocked with Animal-Free Blocker R.T.U. (Vector Laboratories), probed with anti-Luciferase antibodies (Abcam; AB185923, 1:200 dilution) overnight at 4°C, washed with PBS, and incubated with ImmPRESS anti-Rabbit polymer detection reagent (Vector Laboratories) for 30 minutes at room temperature. Visualization was performed by incubation with 3,3′-diaminobenzidine (DAB; Vector Laboratories), tissues were counterstained with Gill No.3 Hematoxylin (Sigma), coverslipped, and imaged on an Olympus IX73 inverted microscope.

Trotter T.N., Wilson A., McBane J., Dagotto C.E., Yang X.Y., Wei J.P., Lei G., Thrash H., Snyder J.C., Lyerly H.K, & Hartman Z.C. (2024). Overcoming Xenoantigen Immunity to Enable Cellular Tracking and Gene Regulation with Immune-competent “NoGlow” Mice. Cancer Research Communications, 4(4), 1050-1062.

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Immunohistochemical Staining of UNC-45A

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Five-micron thick formalin-fixed, paraffin-embedded sections were deparaffinized and rehydrated by sequential washing with xylene, 100% ethanol, 95% ethanol, 80% ethanol, and PBS. For antigen retrieval, slides were immersed in Reveal Decloaker (Biocare Medical, Concord, CA) and steamed for 30 min at 100 degrees C. Endogenous peroxidase activity was blocked with 3% H₂O₂ for 10 min. After washing with PBS, slides were blocked with 10% normal goat serum in PBS for 10 min at room temperature, followed by incubation with rabbit anti-human polyclonal UNC-45A antibody (Proteintech Group Inc) at a concentration of 1:200 in blocking solution overnight at 4℃. After washing twice with PBS, slides were incubated with a biotinylated anti-rabbit secondary antibody conjugated (10 min) and streptavidin/horseradish peroxidase (10 min; Dako), followed by 3,3-diaminobenzidine (Phoenix Biotechnologies) substrate for 3 min. Slides were lightly counterstained with Gill No. 3 hematoxylin (Sigma) for 60 s, dehydrated, and coverslipped.

Habicht J., Mooneyham A., Shetty M., Zhang X., Shridhar V., Winterhoff B., Zhang Y., Cepela J., Starr T., Lou E, & Bazzaro M. (2019). UNC-45A is preferentially expressed in epithelial cells and binds to and co-localizes with interphase MTs. Cancer Biology & Therapy, 20(10), 1304-1313.

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