Assay

4.2.3.5. Antioxidant assays in gastric tissue homogenate 4.2.3.5.1. Catalase activity Catalase activity in the gastric tissue was measured according to the procedures of Sinha (1972). One ml of homogenate solution was taken and mixed with 5 ml of phosphate buffer of pH 7.4. To this 4 ml of 0.2 M H2O2 was added and time noted. Exactly after 180 seconds of adding H2O2, a set of 1ml of reaction mixture from the above was taken and 2 ml dichromate acetic acid was added. It was then kept in a boiling…

cells from a cauliflower were studied with the purpose to separate nuclei and mitochondria content from a solution. To achieve the results, the cauliflower solution had to go through differential centrifugation, microscopy, and succinate dehydrogenase assay. For the fist part of the lab density played a key role in the separation of the cells content because mitochondria and nuclei are very different in size, mitochondria is approximately ten times smaller than nuclei. So it was hypothesized…

(chemoattractants) or unfavourable chemicals (chemorepellents). In particular, sugars and amino acids can attract many species of bacteria via chemotaxis, and the main method used to identify these chemicals is an assay that uses a glass capillary (Abe et al., 2017). In the capillary assay, the outer chamber contains mobile cells while the inner chamber contains the chemical to be tested. If the chemical is a chemoattractant, the cells will migrate in the inner chamber. If it is a…

concentration” (2). IC 50 represents the concentration of the compounds that required for 50% of cell death. The IC 50 assay is used to determine the potency of the drugs which used as treatment of cancers (1). In the previous experiments, 20,000 cells/mL was the optimal number of leukemia cells, which was the concentration of cells given the absorbance values between 0.75 and 1.25 in MTT assay. Moreover, compounds were tested for their possible effect on the reduction of the proliferation of…

Louis, MO), we could determine both mitochondrial outer membrane integrity and cytochrome c oxidase activity. In the colorimetric assay, cytochrome c oxidase via oxidation of ferrocytochrome c to ferricytochrome c could reduce the absorbance at 550 nm. All of the procedures were done according to the manufacturer's method; 20 μg of mitochondrial fraction were applied for each reaction…

Incubation of Macrophage with MSCs 100 µl of MSC suspension (2 × 105 cell/ml) was added to each well of 96-well microplates containing macrophages and incubated for 4 h at 37°C in a moist atmosphere of 5% CO2. 2.5 Phagocytic assay by neutral red The basic phagocytic ability of macrophage (non-opsonic dependent phagocytosis) was measured by neutral red uptake. After MSCs and Macrophages were co-cultured for 2 h, 200 µl neutral red solutions (dissolved in 10 mmol/L PBS with the…

(GE Healthcare) with a flow rate 1 ml/min. Protein and peptides absorbance were detected at 280 nm. 2 ml fractions were collected automatically and tested for antimicrobial activity as previously described in point Bacteriocin detection and activity assay. After checking optimal step for bacteriocin elution (designed as Fraction I), only this was collected for subsequent purification and activity…

together by peptide bonds. As stated by Mbuthia, the Biuret reagent is a solution which aids scientists to conclude if there are any peptide bonds present within a sample, and thus, is used to determine protein existence within a substance. The Biuret assays depends on the presence of 2 or more peptide bonds in proteins. In an alkali environement, when a protein solution is given (Cu2+) Cupric ions, they will be reduced to (Cu+) copper cuprous, thus causing a Cu2+ - copper chelate complex,…

Cytotoxicity assay: Cytotoxic and viability assay of pulchranin A on human cancer cell lines: The isolated compound exhibited a moderate cytotoxic effect on the three tested cell lines: HCT, HepG2 and MCF-7 where the cell viability, measured by SRB assay, and the IC50 was found to be 91, 80, 63 µg/ml respectively (Figure 3). The standard used was doxorubicin, which gave an IC50 value…

lowest TNF alpha value. Also, Figure 8 shows another bar chart of HeLa cell that was stimulated with TNF alpha induced NFkB activation. The differences between stimulated (20ng/ml of TNF alpha) and unstimulated HeLa cell as a result of the luciferase assay…