Ma5 16356

Manufactured by Thermo Fisher Scientific

The MA5-16356 is a laboratory instrument manufactured by Thermo Fisher Scientific. It is designed to perform a specific function within the laboratory environment. The description of the core function of this product is limited to the information available and an unbiased, factual presentation to maintain objectivity.

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Lab products found in correlation

Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Vectastain elite abc hrp kit, by Vector Laboratories (1 mentions) Sc 40, by Santa Cruz Biotechnology (1 mentions) Rm 9106, by Thermo Fisher Scientific (1 mentions) A11055, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) A11034, by Thermo Fisher Scientific (1 mentions) Mouse anti c myc, by Santa Cruz Biotechnology (1 mentions) A11036, by Thermo Fisher Scientific (1 mentions) Sc 15334, by Santa Cruz Biotechnology (1 mentions) Ab769, by Merck Group (1 mentions) Ma5 15257, by Thermo Fisher Scientific (1 mentions) Ma5 15739, by Thermo Fisher Scientific (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions)

3 protocols using ma5 16356

Colon Tissue Microarray Analysis of Cyclin D1 and CD163

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Tissue microarrays for normal and cancerous human colon tissues (sample ID: BC05118d) were purchased from Biomax (Rockville, MD). The tissue microarray samples were dried for 20 minutes at 60 °C, deparaffinized with xylene, and rehydrated. Antigens were retrieved by heating the sections at 95 °C in 0.01 M sodium citrate buffer, pH 6.0. Endogenous peroxidase activity was blocked using 3% hydrogen peroxide. The samples were stained with antibodies against cyclin D1 (1:100, MA5-16356; Thermo Fisher Scientific) and CD163 (1:100, MA5-11458; Thermo Fisher Scientific). Incubation with a biotin-conjugated secondary antibody and visualization were performed using the VECTASTAIN Elite ABC-HRP kit (Vector Laboratories, Burlingame, CA), followed by counterstaining with hematoxylin. The processed slides were analyzed under a light microscope (IX53; Olympus, Tokyo, Japan) using the CellSens Dimension program. Cyclin D1 immunostaining intensity in macrophage-enriched areas of colon tissues was analyzed using ImageJ software (National Institutes of Health, Rockville, MD).

Noh J.Y., Lee I.P., Han N.R., Kim M., Min Y.K., Lee S.Y., Yun S.H., Kim S.I., Park T., Chung H., Park D, & Lee C.H. (2022). Additive Effect of CD73 Inhibitor in Colorectal Cancer Treatment With CDK4/6 Inhibitor Through Regulation of PD-L1. Cellular and Molecular Gastroenterology and Hepatology, 14(4), 769-788.

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Immunostaining of Organoid Cultures

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Immunohistochemistry and immunofluorescence staining of organoids were performed as described previously [21 (link)]. The following primary antibodies were used: rabbit anti-MUC2 (1:200; sc15334, Santa Cruz Biotechnology, Dallas, TX), rabbit anti-TFF3 (5 μg/ml; ab101099, Abcam, Cambridge, MA), rabbit anti-Ki67 (1:150; RM-9106, Thermo Fisher, Carlsbad, CA), rabbit anti-β-catenin (1:200; #9582, Cell Signaling Technology), mouse anti-C-myc (1:200; sc40, Santa Cruz Biotechnology), and rabbit anti-cyclin D1 (1:150; MA5–16356, Thermo Fisher Scientific).
For immunofluorescence staining, primary antibodies were rabbit anti-cleaved caspase-3 (1:400; #9661, Cell Signaling Technology), rabbit anti-laminin 332 (1:500; kind gift from Peter Marinkovich, Stanford University, Stanford, CA), and goat anti-collagen IV (1:100; AB769, Millipore). Secondary antibodies were AlexaFluor 568 goat anti-rabbit IgG (1:500; A-11036, Invitrogen), AlexaFluor 488 donkey anti-goat IgG (1:500; A-11055, Invitrogen), and AlexaFluor 488 goat anti-rabbit IgG (1:500; A-11034, Invitrogen). Nuclei were counterstained with Hoechst 33342 (Thermo-Fisher Scientific).

Liu X., Cheng Y., Abraham J.M., Wang Z., Wang Z., Ke X., Yan R., Shin E.J., Ngamruengphong S., Khashab M., Zhang G., McNamara G., Ewald A.J., Lin D., Liu Z, & Meltzer S.J. (2018). Modeling Wnt signaling by CRISPR-Cas9 genome editing recapitulates neoplasia in human Barrett epithelial organoids. Cancer letters, 436, 109-118.

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Western Blot Analysis of Cell Signaling Proteins

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The cells were lysed in a radioimmunoprecipitation assay buffer containing protease inhibitor cocktail (Roche, Mannheim, Germany). Pierce Coomassie Protein Assay (Bradford, Thermo Fisher Scientific, Waltham, MA, USA) was applied to measure the concentrations of protein extracts. Aliquots of protein extracts (40 µg/lane) were subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to phosphatidylcholine membranes (Sartorius, Göttingen, Germany). The membranes were blocked using skim milk and separately incubated overnight at 4 °C with primary antibodies against RFP (1:1000, MA5-15257, Thermo Fisher Scientific), Cre (1:1000, ab188568, Abcam), CDK4 (1:1000, MA5-12984, Thermo Fisher Scientific), cyclin D1 (1:200, MA5-16356, Thermo Fisher Scientific), cyclin A2 (1:500, 18202-1-AP, Proteintech), cyclin B1 (1:500, 55004-1-AP, Proteintech), and β-actin (1:5000, MA5-15739, Thermo Fisher Scientific). Subsequently, the membranes were probed using HRP-conjugated rabbit/mouse anti-IgG for 1 h at room temperature. Finally, protein bands were detected through enhanced chemiluminescence (ECL; PerkinElmer Life Science, Hopkinton, MA, USA) using a UVP Biospectrum (UVP, LLC Upland, CA, USA).

Tseng H.C., Wu M.R., Lee C.H, & Hsiao J.K. (2022). Differentiation Capacity of Bone Marrow-Derived Rat Mesenchymal Stem Cells from DsRed and Cre Transgenic Cre/loxP Models. Cells, 11(17), 2769.

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