BALF was clarified by centrifugation, aliquoted and stored at −80 °C for later analysis. Cytokine levels were measured in BALF using ELISA kits from eBioscience (San Diego, CA) and R&D Systems (Minneapolis, MN). Myeloperoxidase (MPO) activity in BALF samples was measured using the Myeloperoxidase activity fluorometric assay kit from BioVision (Mountain View, CA). Eosinophil peroxidase (EPO) in BALF samples was measured using an Eosinophil peroxidase ELISA kit (Cusabio Biotech, Wuhan, China). Soluble collagen levels in BALF were measured by adding 200 µL of 0.1 % Direct Red 80 in picric acid (Sigma Aldrich, St. Louis, MO) to 50 µL of lavage fluid and incubating for 60 min at 37 °C. The samples were centrifuged and the pellet was washed with 100 % ethanol. The pellet was resuspended in 200 µl 0.5 M NaOH, incubated for 30 min at 37 °C and the absorbance was read at 540 nm.
Direct red 80 in picric acid
Manufactured by Merck Group
Direct Red 80 in picric acid is a laboratory reagent used as a staining dye. It is a red-colored powder that is soluble in water and alcohol. The product is typically used in various laboratory applications, including histology, cytology, and microscopy, to stain biological samples for visualization and analysis purposes. The core function of this product is to provide a staining solution for coloring and highlighting specific structures or components within a sample.
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Lab products found in correlation
2 protocols using direct red 80 in picric acid
1
Quantification of Inflammatory Markers in BALF
2
Quantifying Collagen Formation in Preosteoblasts
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To determine the formation of collagenous matrix, collagen content in the preosteoblastic culture was stained with Direct Red (Sigma-Aldrich Co.) for 3, 9, 15, and 21 days. MC3T3-E1 cells were plated in a 48-well plate, and after overnight incubation, the cells were treated with AnTT. At the end of each time point, the cells were washed with PBS, and 0.1% Direct Red 80 in picric acid (Sigma-Aldrich Co.) was added into the wells. The plates were then incubated for 1 h at 37°C. The cells were washed with 10 mM hydrochloric acid (Fisher, Hampton, VA, USA), and the staining was visualized under the microscope EVOS Cell Imaging System. The bound collagen was dissolved with 0.1 M sodium hydroxide, and the absorbance was measured at 450 mm using a microplate reader.
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