With prior IACUC approval from the Indiana University School of Medicine (#10797), twenty five 11 week old female C57BL/6 mice (Envigo, Indianapolis, IN) were sacrificed via CO2 inhalation in accordance with the National Institutes of Health guide for the care and use of Laboratory animals, at which time the left and right tibiae were harvested and stripped of soft tissue. The tibiae were randomly separated into 2 preservation groups (both tibiae from each animal were kept in the same group). Each tibia was stored individually in a microcentrifuge tube under one of the following conditions: wrapped in gauze soaked in phosphate buffered saline (Gibco PBS pH 7.4, Thermo Fisher Scientific, Waltham, MA; PBS) at −20 °C (n = 13) or submerged in 70% ethanol at 4 °C (n = 12). In total, the bones were stored for 7 weeks before beginning the ribosylation experiment.
Gibco pbs ph 7
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
Gibco PBS pH 7.4 is a sterile, ready-to-use phosphate-buffered saline solution. It maintains a physiological pH of 7.4 and is commonly used as a general-purpose buffer in various laboratory applications.
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Lab products found in correlation
2 protocols using gibco pbs ph 7
1
Murine Tibia Preservation Techniques
2
Acai Antioxidant Reducing Capacity Assay
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The ability to reduce ferric ions (Fe3+) to ferrous ions (Fe2+) was used to estimate the reducing capacity of acai extracts. The acai extract concentrations were assessed over a concentration range of 0.001–8000 µg/mL. Each assay data point contained 4 µL of each acai extract, 400 µL of phosphate buffer (Gibco™ PBS, pH 7.4, ThermoFisher, Stafford, UK), and 250 µL of 1% potassium ferricyanide. After the incubation of the mixture at 50 °C for 20 min, 250 µL of 10% trichloroacetic acid was added. The samples were centrifuged at 3000 rpm for 10 min. Then, 100 µL of the supernatant was transferred to a 96-well microtiter plate and mixed with 100 µL of double-distilled water and 20 µL of freshly prepared (0.1%) ferric chloride (FeCl3) solution. Then, the formation of Perl’s Prussian blue was read at 700 nm, according to Nwidu et al. (2018) [82 (link)] using a Varioskan™ LUX multimode microplate reader (ThermoFisher, Stafford, UK). The positive control was L-ascorbic acid.
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