Ht501320

Manufactured by Merck Group

Sourced in United States, Germany

The HT501320 is a laboratory equipment product. It is designed for general laboratory use, but its specific core function is not available in the provided information. A more detailed and unbiased description cannot be provided without extrapolation.

Automatically generated - may contain errors

Lab products found in correlation

Inverted microscope, by Leica (2 mentions) Ab15580, by Abcam (2 mentions) A5486, by Merck Group (1 mentions) Sm2010r, by Leica (1 mentions) Ab152, by Merck Group (1 mentions) Cy3 conjugated donkey anti rabbit, by Jackson ImmunoResearch (1 mentions) Dfc365fx, by Leica (1 mentions) Dm5500 microscope, by Leica (1 mentions) D3571, by Thermo Fisher Scientific (1 mentions) Permafluor mounting medium, by Thermo Fisher Scientific (1 mentions) Zen 2, by Zeiss (1 mentions) A412 4, by Thermo Fisher Scientific (1 mentions) Histocore multicut, by Leica (1 mentions) Trichrome stain masson kit, by Merck Group (1 mentions) Bx43 light microscope, by Olympus (1 mentions)

14 protocols using ht501320

Colitis and Colorectal Cancer Induction in Mice

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Male and female littermate WT, Ku70^+/−, and Ku70^−/− mice of 8 to 10 weeks were injected intraperitoneally with 10 mg of AOM (A5486, Sigma-Aldrich) per kilogram body weight. Five days later, 1.5% DSS (160110, MP Biomedicals) was given in the drinking water for 6 days, followed by regular drinking water for 2 weeks. This cycle was repeated twice with 1.5% DSS. Mice were ethically culled on day 80, colon tissues were harvested, and the colon length was measured. Tumor numbers were enumerated, and the colon tissue was stored at −80°C or fixed in 10% neutral-buffered formalin (NBF) (HT501320, Sigma-Aldrich) for further analysis.
For colitis induction, mice were injected with AOM and, after 5 days, fed with 1.5% DSS for 6 days. Mice were then given regular water for 3 days and were ethically culled on day 14. On day 14, colon tissues were harvested, colon length was measured, and the tissue was stored at −80°C or fixed in 10% NBF for further analysis.

Pandey A., Shen C., Feng S., Enosi Tuipulotu D., Ngo C., Liu C., Kurera M., Mathur A., Venkataraman S., Zhang J., Talaulikar D., Song R., Wong J.J., Teoh N., Kaakoush N.O, & Man S.M. (2024). Ku70 senses cytosolic DNA and assembles a tumor-suppressive signalosome. Science Advances, 10(4), eadh3409.

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Immunohistochemical Analysis of Neuronal Markers

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Following completion of experiments, rats were anesthetized with sodium pentobarbital (100 mg/kg) and transcardially perfused with 0.01 M PBS (phosphate-buffered saline) followed by 10% buffered formalin solution (HT501320, Sigma Aldrich). Brains were removed and stored in formalin with 20% sucrose. All brains were sectioned at 30 μm on a freezing stage microtome (SM2010R, Leica Biosystems). Sections were collected and processed to label for GFP (as an indicator of GCaMP6f or dLight1.2 expression) and/or TH via immunohistochemistry. Antibodies were incubated at 4 °C (washes and other steps at room temperature). Tissues were permeabilized in 0.3% Triton-X 100 for 30 min and blocked in 2% normal donkey serum for 30 min. Sections were incubated in rabbit anti-TH (AB152, Sigma Aldrich) and/or chicken anti-GFP (AB13907, Abcam) antibodies overnight (∼18 h). After KPBS (potassium phosphate-buffered saline) washes (eight changes, 10 min each), secondary antibody (Cy3 conjugated donkey anti-rabbit and AF488 conjugated donkey anti-chicken; Jackson Immunoresearch) was applied and sections were incubated overnight. Sections were then mounted onto glass slides, air dried, and coverslipped with 50% glycerol in KPBS mountant. Only data from subjects with GFP and mCherry expression and correct fiber placement were included in statistical analyses.

Hsu T.M., Bazzino P., Hurh S.J., Konanur V.R., Roitman J.D, & Roitman M.F. (2020). Thirst recruits phasic dopamine signaling through subfornical organ neurons. Proceedings of the National Academy of Sciences of the United States of America, 117(48), 30744-30754.

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Histopathological Lung Tissue Analysis

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Lung tissues were fixed with 10% neutral buffered formaldehyde (HT501320; Sigma-Aldrich) overnight, and dehydrated, embedded in paraffin. Then the tissues were sectioned for staining with hematoxylin and eosin (H&E) for histopathology.

Liu L., Lei Y., Chen W., Zhou Q., Zheng Z., Zeng G., Liu W., Feng P., Zhang Z., Yu L, & Chen L. (2022). In vivo genome-wide CRISPR screening identifies ZNF24 as a negative NF-κB modulator in lung cancer. Cell & Bioscience, 12, 193.

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Comprehensive Pathological Examination of Mice

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Mice were euthanized by CO2 asphyxiation followed by complete pathological examination including necropsy with dissection and histological analysis of the following organ and tissues: head in toto, skin (including site of transplantation and auricles), salivary glands, larynx and thyroids, trachea, lungs, heart, mediastinum, spleen, liver, pancreas, kidneys, esophagus and stomach, small intestine, large intestine, urogenital tract, cervical and tracheobronchial lymph nodes, sternum. Samples were immersion-fixed in 10% neutral buffered formalin (Sigma-Aldrich #HT501320), routinely processed for paraffin embedding, sectioned at 5 μm and stained with Hematoxylin (Diapath #C0302) and Eosin (Diapath #C0362). Serial sections obtained from representative samples were also stained by means of immunohistochemistry (IHC) or in situ hybridization (ISH) as detailed in the S2 Table. Head and sternum were decalcified in a 14% solution of Tetrasodium EDTA (VWR BDH Prolabo #20299291) for 15 days before processing and paraffin embedding.

Radaelli E., Hermans E., Omodho L., Francis A., Vander Borght S., Marine J.C., van den Oord J, & Amant F. (2015). Spontaneous Post-Transplant Disorders in NOD.Cg- Prkdcscid Il2rgtm1Sug/JicTac (NOG) Mice Engrafted with Patient-Derived Metastatic Melanomas. PLoS ONE, 10(5), e0124974.

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Tissue Imaging and Colocalization Protocol

Cited 1 time

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Conducted as described previously^{16 (link)}. Briefly, tissues for imaging were fixed overnight in (mouse) or for 7 days (NHP) in 10% neutral buffered formalin (Sigma, HT501320), and then transferred to PBS. All tissues were paraffin imbedded and sliced into 4 μM sections. Sections were stained with DAPI and imaged on a Leica inverted microscope. Staining for colocalization of Di-siRNA with neuronal (NueN) and glial cells (GFAP) was performed as described previously^{50 (link)}.

Alterman J.F., Godinho B.M., Hassler M.R., Ferguson C.M., Echeverria D., Sapp E., Haraszti R.A., Coles A.H., Conroy F., Miller R., Roux L., Yan P., Knox E.G., Turanov A.A., King R.M., Gernoux G., Mueller C., Gray-Edwards H.L., Moser R.P., Bishop N.C., Jaber S.M., Gounis M.J., Sena-Esteves M., Pai A.A., DiFiglia M., Aronin N, & Khvorova A. (2019). A divalent siRNA chemical scaffold for potent and sustained modulation of gene expression throughout the central nervous system. Nature biotechnology, 37(8), 884-894.

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Tissue Imaging and Colocalization Protocol

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Biodistribution of Labeled siRNA in Mice

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C57BL/6 mice were injected intravenously (IV, via tail vein) or subcutaneously (SC, interscapular, between shoulders) with 10 mg/kg of Cy3-labeled hsiRNA (Cy3-hsiRNA^sFLT1 or Cy3-FM-hsiRNA^sFLT1). After 24 h mice were sacrificed and liver, kidney, spleen, intestine, muscle (quadriceps), pancreas, lung tissues, as well as fat, skin and tail from the site of injection, were removed and fixed in 10% formalin (Sigma, #HT501320) overnight at 4°C. Fixed tissues were embedded in paraffin, and sliced into 4 μm sections that were mounted on glass slides. Sections were deparaffinized by incubating in Xylene twice for 8 min. Sections were rehydrated in serial ethanol solutions (100%, 95%, 80% dilutions in water) for 4 min each, then washed twice for two minutes with PBS. Washed slides were incubated with 0.25 μg/ml DAPI (Molecular Probes, #D3571) in PBS for 1 min then washed twice for 2 min with PBS. Slides were mounted with PermaFluor mounting medium (Thermo, #TA030FM) and coverslips and dried overnight before imaging on a Leica DM5500 microscope fitted with a DFC365 FX fluorescence camera.

Hassler M.R., Turanov A.A., Alterman J.F., Haraszti R.A., Coles A.H., Osborn M.F., Echeverria D., Nikan M., Salomon W.E., Roux L., Godinho B.M., Davis S.M., Morrissey D.V., Zamore P.D., Karumanchi S.A., Moore M.J., Aronin N, & Khvorova A. (2018). Comparison of partially and fully chemically-modified siRNA in conjugate-mediated delivery in vivo. Nucleic Acids Research, 46(5), 2185-2196.

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Histological analysis of xenograft tissues

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After euthanasia, the xenograft or lung tissues were dissected from mice and fixed in 10% neutral buffered formalin solution (HT501320; Sigma‐Aldrich) overnight. The tissues were dehydrated, paraffin embedded, and sectioned for staining with hematoxylin and eosin or anti‐Ki67 (Rb mAb, ab15580, Abcam) for immunohistochemistry.

Zhou Q., Liang J., Yang T., Liu J., Li B., Li Y., Fan Z., Wang W., Chen W., Yuan S., Xu M., Xu Q., Luan Z., Xia Z., Zhou P., Huang Y, & Chen L. (2021). Carfilzomib modulates tumor microenvironment to potentiate immune checkpoint therapy for cancer. EMBO Molecular Medicine, 14(1), e14502.

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Quantifying Hepatic Ischemia-Reperfusion Injury

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Liver samples, that were preserved in a 10% formalin solution (HT501320, Sigma-Aldrich, St Louis, MO), were stained using hematoxylin and eosin, and a pathologist blinded to the experimental allocation assessed the Suzuki score, which represents the degree of hepatic IRI. The Suzuki score was derived from the mean value of scores (0–4) for 7 criteria to assess the degree of hepatic IRI: inflammation, necrosis, overall degree of IRI, congestion, ballooning, steatosis, and cholestasis.^{24 (link)}

Kim J., Hagen C.E., Kumar S.N., Park J.I., Zimmerman M.A, & Hong J.C. (2020). Anticholestatic Effect of Bardoxolone Methyl on Hepatic Ischemia-reperfusion Injury in Rats. Transplantation Direct, 6(8), e584.

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Visualizing Brain Vasculature through Latex Perfusion

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To visualize the brain vasculature, systemic latex dye perfusion was performed. Blue latex dye (BR80B, Connecticut Valley Biological, Southampton, MA, USA) was injected into the left ventricle of the heart. Brains were collected and fixed in 10% neutral buffered formalin solution (HT501320, Sigma-Aldrich, St. Louis, MO, USA). The brains were sequentially dehydrated with a series of methanol (A412-4, Fisher Scientific, Hampton, NH, USA) solutions (50%, 75%, 95% and 100% in each 24 h), and cleared in benzyl benzoate (AC105862500, ACROS organic, Fair Lawn, NJ, USA) and benzyl alcohol (AC148390010, ACROS organic), (1:1 ratio). Each single dysplastic lesion image was captured under 30× magnification using a light microscope (Zeiss SteREO Discovery. V12, Jena, Germany). To measure the dysplastic vessel size, the regions of interest (ROIs) were drawn with a spline contour and automatically quantified by a blinded expert using Zen 2.3 software (Zeiss).

Park E.S., Kim S., Yao D.C., Savarraj J.P., Choi H.A., Chen P.R, & Kim E. (2022). Soluble Endoglin Stimulates Inflammatory and Angiogenic Responses in Microglia That Are Associated with Endothelial Dysfunction. International Journal of Molecular Sciences, 23(3), 1225.

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