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TPC were determined also by FTIR method, either using the band intensity at 1742 cm–1 or from the area between the region 950 1900 cm–1, with reference to the calibration curve obtained with pure GAE (range of concentrations 2.5 30 ng/mL M) [35].
The result for each sample was graded as recommended using the band intensity from 0 to 4+.
The H3 K9 sensor developed in the present study is expressed approximately at 15 μM in transfected cells (calculated using the band intensity of Western blot analysis of sensor proteins by FLAG antibody by comparing with tubulin).
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For quantification of band intensity, the gel analysis function of ImageJ (National Institute of Health, USA) was used and the band intensity was standardised to β-tubulin.
The Alphalmager Gel Imaging System (Alpha Innotech, CA) was used to quantify the expression of proteins and the Image J processing software was used to quantify the band intensity.
This programme was also used to analyse the band intensity of Northern and Southern blots.
the blots were visualized using WesternBreeze Kit (WB7105, Invitrogen Life Technologies, CA, USA), and further quantified using Quantity One Software Bio-Radd Laboratories, USA) by measuring the band intensity for each group and normalizing to β-Tubulin and GAPDH as internal control.
Reaction were carried out at the following conditions: hot start at 95°C for 12 min, followed by 32 cycles of 94°C for 30 s, 58°C for 30 s and 72°C for 30 s, and final extension at 72°C for 10 min. Semi-quantitative analysis was performed by determining the band intensity using AlphaEaseFC software (Alpha Innotech).
In case the alcoholic solvent is used, the excitonic band intensity increases approximately 30 times.
The relative expression of genes was determined by measuring the band intensity and using cyclophilin A as housekeeping [67].
Western blotting bands were quantified using Odyssey v3.0 software by measuring the band intensity (area×absorbance) for each group and normalizing to GAPDH band as an internal control.
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