5 lox

Manufactured by Santa Cruz Biotechnology

Sourced in United States

5-LOX is an enzyme that catalyzes the conversion of arachidonic acid into leukotrienes. It plays a key role in the inflammatory response.

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3 protocols using 5 lox

Immunohistochemical Profiling of Stem Cell Markers

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5 μm fixed sections were incubated with primary antibodies in a hybridization chamber for 1 h at room temperature or overnight at 4°C. The primary antibodies used were DclK1, COX-2, 5-LOX, Ki67, proliferating cell nuclear antigen (PCNA), CD133, CD44, Lgr5, Annexin V and β-catenin procured from Santa Cruz/Abgent/Abcam/Abcam/Cell Signaling. Following primary antibody, sections were incubated for 1 h with anti-mouse/anti-rabbit/anti-goat secondary antibody, then visualized with diaminobenzidine (DAB) and counterstained with H&E for IHC or with DAPI for immunohistofluorescence (IHF). Slides were observed under an Olympus microscope 1X701 and digital computer images were recorded with an Olympus DP70 camera.

Mohammed A., Janakiram N.B., Madka V., Brewer M., Ritchie R.L., Lightfoot S., Kumar G., Sadeghi M., Patlolla J.M., Yamada H.Y., Cruz-Monserrate Z., May R., Houchen C.W., Steele V.E, & Rao C.V. (2015). Targeting pancreatitis blocks tumor-initiating stem cells and pancreatic cancer progression. Oncotarget, 6(17), 15524-15539.

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Isoorientin Extraction and Characterization

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MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), trypsin-EDTA, Tris, ethylenediaminetetraacetic acid (EDTA), diethyldithiocarbamate (DDC), Tween-20, hematin, glycerol, phenol, and ammonium sulphate were purchased from Sigma Chemical Co. (St. Louis, MO). Celecoxib was a generous gift from Unichem Laboratories (Mumbai, India). Carrageenan was purchased from Sigma-Aldrich (St. Louis, USA). Polyclonal antibodies to COX-2, IL-6, 5-LOX, IL-1β, and TNF-α were purchased from Santa Cruz Biotechnology (California, USA). iNOS antibody was from Thermo Fisher Scientific Inc. All other chemicals and solvents were of analytical grade and purchased from authorized standard companies. Isoorientin was isolated from the methanolic extracts of tubers of P. tuberosa [13 (link)].

Anilkumar K., Reddy G.V., Azad R., Yarla N.S., Dharmapuri G., Srivastava A., Kamal M.A, & Pallu R. (2017). Evaluation of Anti-Inflammatory Properties of Isoorientin Isolated from Tubers of Pueraria tuberosa. Oxidative Medicine and Cellular Longevity, 2017, 5498054.

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Investigating MCF-7 Cell Line Metastasis

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The MCF-7 cell line was obtained from the American Type Culture Collection (ATCC, USA). Female BALB/c nude mice (18–22 g, 5–6 weeks old) were obtained from the Experimental Animal Center of Nanchang University. BML-111 (Cayman Chemical, USA) was stored at −80°C before it was diluted in a culture medium without serum. Anti-human/mouse antibodies (Santa Cruz Biotechnology, USA), including 5-LOX, 12-LOX, 15-LOX, vimentin, E-cadherin, MMP-2, MMP-9, and β-Actin, were used in this study. CoCl₂ and butoxycarbonyl-Phe-Leu-Phe-Leu-Phe (Boc-2) were purchased from Sigma–Aldrich (USA) and GenScript Corporation (USA). A bicinchoninic acid protein (BCA) kit was used to determine the protein concentration of cell lysates (Pierce, USA). Cell culture medium and fetal bovine serum (FBS) were purchased from Gibco (Australia) and Gibco BRL (Switzerland), respectively. Hoechst 33258 stain was purchased from Novus (Burlington, MA, USA).

Xu F., Zhou X., Lin L., Xu J., Feng Y., He Y, & Hao H. (2023). BML-111, the agonist of lipoxin A4, suppresses epithelial-mesenchymal transition and migration of MCF-7 cells via regulating the lipoxygenase pathway. International Journal of Immunopathology and Pharmacology, 37, 03946320231223826.

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