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Propidium Iodide Staining Protocol Flow Cytometry
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Analysis Of Bf Viability By Flow Cytometry. A) On The Left Are…
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From Muhammad Din Ibrahim 1, Saie Brindha Kntaya 1, Nooraini Mohd Ain 1, Renato Iori 2, Costas Ioannides 3 and Ahmad Faizal Abdull Razis 4, 5, 6, *
CREA Advice on Agricultural Research and Agricultural Economics Research, Center for Agricultural and Climate Research (CREA-AA), By Corticella 133, 40128 Bologna, Italy
Food Safety and Integrity Laboratory, Tropical Agriculture and Food Security Center, Putra Malaysia University, 43400 UPM Serdang, Selangor, Malaysia
Propidium Iodide (pi) Staining Solution (50μg/ml) E Ck A161 Manufacturer
Glucoraphasatin (GR), a glucosinolate that is abundant in plants of the Brassicaceae family, is made by hydrolyzed to rafasatin, which is considered responsible for its own chemopreventive activity; However, the mechanisms underlying them have not been investigated, especially in cell lines. The aim of this study was to determine the cytotoxicity of rafasatin, and to evaluate its potential to induce apoptosis and cell proliferation in breast adenocarcinoma MCF-7 cells. Cytotoxicity was determined after cell injection with glucoraphasatin or rafasatin (0–100 μM), for 24, 48 and 72 hours. GRH did not show cytotoxicity as compared to 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT). When mirosinase was added to the preparation system to convert GRA to rafasatin, cytotoxicity appeared. Exhaustion of cells to rafasatin resulted in apoptosis, as exemplified by cell degeneration, membrane inflammation, chromatin inflammation and nuclear disruption. Furthermore, using annexin V-FITC assay, rafasatin induces apoptosis, as evidenced by changes in cell division, at different stages of apoptosis; In addition, cleavage resulted in the capture of MCF-7 Cells in G
+ B. Finally, rafasatin showed potential immunosuppressive effect on breast cancer adenocarcinoma (MCF-7), by inducing apoptosis and arresting cell cycle.
Glucoraphasatin; Rafasatin; Glucosinolates; Isothiocyanates; Human breast cancer adenocarcinoma cell line; Cytotoxicity; Apoptosis; glucoraphasatin cell cycle; Rafasatin; Glucosinolates; Isothiocyanates; Human breast cancer adenocarcinoma cell line; Cytotoxicity; Apoptosis; Cell cycle
Cancer fails to balance the growth and diversity of organisms, and has been a major cause of death worldwide. Although chemo- and radio-therapy are now frequently used in the treatment of cancer , they are associated with serious side effects, as a result of damage to the tissues surrounding the tumor . Furthermore, tumors may show resistance to these treatments . Breast cancer is a common disease among Malaysian women and around the world, with around 150-200 new cases being diagnosed each year at the Breast Hospital at Kuala Lumpur Hospital alone. It is estimated that 1 in 20 women will be diagnosed with breast cancer [4, 5]. Cancer accounted for 16.5% of all cancers registered with the Malaysian Cancer Register in 2006, and 34.4% of breast cancer patients were between 15-49 years of age .
Facs Plots Showing: A) The Exclusion Of Non Viable Propidium Iodide (pi…
Over the past several decades, dietary importance has been regulated in the etiology of human cancer . Many phytochemicals, including ascorbic acid and phenolics, such as anthocyanins and flavonoids, have antioxidant activity and are considered to be important contributors to the beneficial effects associated with eating a rich vegetable diet. . Broccoli (Brassica oleracea), radish (Raphanus sativus L.), kale, and cabbage (B. oleracea L.) are examples of brassica vegetables rich in antioxidants, as well as glucosinolates, which have attracted a lot of attention in the ‘ in recent years. Time to Learn. Age, due to the significant anti-carcinogenic activity of isothiocyanates, their main hydrolysis products [8, 9].
In a cohort study of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis. acute myocardial infarction acute myocardial infarction acute myocardial infarction acute myocardial disease n) of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis of the pathogenesis. acute myocardial infarction acute myocardial infarction acute myocardial infarction acute myocardial infarction. acute myocardial infarction acute myocardial infarction acute myocardial infarction acute myocardial infarction acute myocardial infarction Infectious Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases Diseases The hemopreventive properties of these vegetables are recognized due to the presence of glucosinolates, a class of sulfur-containing glycosides that are present in high concentrations . Glucosinolates (GLS) are the second metabolites of brassica vegetables that are related to their health-promoting benefits, because their hydrolase products, isothiocyanates, have chemopreventive and antioxidant properties .
4-Methylsulfanil-3-butenyl glucosinolate, also called GR, is the glucosinolate, the main source being Rafanus sativus (Cayware daikon), a white radish consumed in Japan and growing in Europe and 13 US. ], 14]. Raphanus sativus L. radishes have recently attracted the attention of the scientific community due to their health-promoting properties. The focus is, in particular, on isothiocyanate rafasatin (Fig. 1), which is released by mirosinase hydrolysis, on chewing, cutting or other breakdown of fresh (unripe) vegetable extracts . Along with contributing to the taste of radish radish, isothiocyanates are potent direct antioxidants that have a chemopreventive immune function and potentially therapeutic activity .
The ability to induce apoptosis is an important component of a candidate’s cancer treatment. Several methods have been developed to evaluate the potential of chemical agents to enhance apoptosis, including biological studies, bacteriological analysis, ELISA, ongoing cytometry studies and others, for which biological research has become more reliable . As previous studies have shown that isothiocyanates exhibit their chemopreventive activity, at least in part, by regulating the apoptosis process and cell proliferation [18, 19, 20, 21], current studies have been conducted. to establish the possibility and the possibility of GR. Rafasatin product, to initiate apoptosis and capture cell cycle in breast cancer adenocarcinoma (MCF-7).
A Stable Propidium Iodide Staining Procedure For Flow Cytometry.
In this study, the potential of MCF-7 cells was investigated after the appearance of GRH and rafasatin. At the analyzed analyzes (1-100 μM), GRH had no effect on MCF-7 cells (Table 1). In contrast, its anti-inflammatory, isothiocyanate rafasatin was cytotoxic, after processing for 24, 48 and 72 hours. Paclitaxel (0–25 nm) acted as an effective control and was highly cytotoxic to cells.
2.2. Morphological study of apoptosis using end deoxynucleotidyl transferase DUTP nick end labeling (TUNEL) assay, acridine orange / propidium iodide (AO / PI) stain, and 4 ′, 6-diamidino-2-phenylpindulin.
In the anatomical analysis, TUNEL Assay, AO / PI staining and DAPI were used to assess climate change in MCF-7 cells after the appearance of rafasatin at different time points, i.e., 24, 48 and 72 hours. In the TUNEL test the untreated cells (Fig. 2A) are healthy, have a round shape and a negative TUNEL. After 24 hours of treatment (Fig. 2b), the cells began to turn a dark brown color indicating that some cells had acquired apoptosis in the early stages. After treatment for 48 h (Fig. 2C), the number of dark-colored cells increased; About 50% of the cell population undergoes apoptosis, which results in nuclear DNA division. Finally, treated 72 T cells showed the highest number of apoptotic cells, covering about 70% of the cell population (Fig. 2D).
In the AO / P spot, the undifferentiated cells (Fig. 3a) did not show any physiological changes, and only a greenish, round-shaped, cells were observed. can be used, which indicate signs of apoptosis. However, apoptosis became apparent after 24 h (Fig. 3 B) treatment with rafasatin, where early apoptosis with central dissociation and chromatin condensation features remained visible, and round-shaped cells also began to shrink. After 48 hours of treatment (Fig. 3C), apoptosis was evident, with membrane inflammation and chromatin condensation visible. The lesion surrounding the single cell suggests that apoptosis has developed. In addition, some necrotic cells are clearly visible, along with red cells. Cells exposed to radiation for 72 hours (3D image) showed the greatest apoptosis, including membrane inflammation, late apoptosis, secondary necrosis, and apoptotic formation. Furthermore, in the late stages of apoptosis, changes such as the presence of red-orange color, due to the binding of AO to the removed DNA, were observed after 72 h of cell treatment.
Propidium Iodide Staining Underestimates Viability Of Adherent Bacterial Cells
As shown in Figure 4, the DAPI stain shows that the MCF-7 molecule treated with rafasatin, for different periods (24, 48 and 72 hours), induces apoptotic cell proliferation, including Nuclear dissociation, formation of micronucleus. , And chromatin condensation, compared to process molecules that retain a round shape. In contrast, treated cells exhibited morphological features of
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