The quantity of the extracted DNA was evaluated using the absorbance at 260 nm, which was converted into ng/μL of double-stranded DNA using the established conversion factor of 50 ng/μL for one optical density unit at 260 nm (Sambrook et al. 1989).
All extractions were assessed in parallel and performed in triplicate, and gDNA recovery was evaluated using UV absorbance (420 ng corresponding to total recovery from 70,000 cells).
The formulation was evaluated using an oxygen radical absorbance capacity assay, in which a fluorescent probe's loss of fluorescent intensity is monitored in the presence of oxygen radicals.
The antioxidant in vitro activity of the extracts obtained from selected cereal products was evaluated using an Oxygen Radical Absorbance Capacity (ORAC) procedure (Brunswick Laboratories, Wareham, MA, USA).
TNF- α and IL-6 were measured using high sensitivity ELISA assays (Thermo Scientific kits, Rockford, IL), with range of 15.6 1000 pg/mL and 10.24 400 pg/mL, respectively, according to instructions provided by the manufacturer and evaluated using an iMark Microplate Absorbance Reader (Bio-Rad Laboratories HercUSAs, CA, USA).
To ensure potent biological activity of the ethanolic extract of Padma 28, its antioxidant capacity was evaluated using the Oxygen Radical Absorbance Capacity (ORAC) assay, which measures the direct capacity of chain-breaking antioxidants based on the hydrogen atom transfer mechanism in a cell-free system.
In addition, alteration of secondary structure of HSA in the presence of butoben was evaluated using the data obtained from UV visible absorbance, CD and FT-IR spectroscopies.
The absorbance of formazan was evaluated using Bio-Tek Synergy HT plate reader (Bio-Tek Instruments Inc., USat at 570 nm.
Cell viability was evaluated using a CCK-8 assay by measuring absorbance.
