Leaves were taken at 60 days after transplanting to assess the chlorophyll and carotenoids content, as described by Witham et al. [32 ]. Briefly, one gram of leaf tissue was crushed in 80% pre-chilled acetone and the volume was made up to 100 mL with pre-chilled acetone. The absorbance of the supernatant was recorded at 452, 663, and 645 nm using a UV–vis 1700 spectrophotometer, Shimadzu, Japan. The amount of chlorophyll and carotenoids present (mg/g) in leaf tissue was calculated by the formula given in Sadashivam and Manickam [33 ].
Uv vis 1700 spectrophotometer
Manufactured by Shimadzu
Sourced in Japan
The UV–vis 1700 spectrophotometer is a compact and versatile instrument designed for absorption and transmission measurements in the ultraviolet and visible light regions of the electromagnetic spectrum. It features a double-beam optical system and a monochromator to provide accurate and reliable data.
Automatically generated - may contain errors
Lab products found in correlation
P53 pan elisa kit, by Roche (3 mentions)
Originpro 8, by OriginLab (3 mentions)
Uv 2550 spectrophotometer, by Shimadzu (1 mentions)
No 2 filter paper, by Cytiva (1 mentions)
Atr pro one, by Jasco (1 mentions)
8400 s fourier transform infrared spectrometer ft ir, by Shimadzu (1 mentions)
Quercetin, by Merck Group (1 mentions)
Ta xt plus texture analyzer, by Stable Micro Systems (1 mentions)
Rotary evaporator, by Heidolph (1 mentions)
Whatman no 2 filter paper, by Cytiva (1 mentions)
17 protocols using uv vis 1700 spectrophotometer
1
Measuring Leaf Pigments: Chlorophyll and Carotenoids
2
Quantifying p53 Protein in Cancer Cell Lines
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The p53 protein produced by transfection of HeLa or N2a cancer cells with the developed ternary systems has been determined by p53 pan ELISA kit (Roche Applied Science, Penzberg, Germany), following the procedure described by the manufacturer. This method is based on a sandwich ELISA principle, where the investigated sample reacts with the capture antibody and peroxidase labelled detection antibody forming a stable complex. The formed colored compound is quantified by spectrophotometry and is proportional to the p53 concentration. Briefly, transfected cell lysates were prepared from detergent lysis in 1% Triton X-100 and 0.1% SDS in PBS, pH 7.4 and proteinase inhibitor cocktail. The biotin-labeled capture antibody is prebound to the streptavidin-coated microtiter plate. During a single incubation step, the p53-containing sample reacts with the capture antibody and, subsequent to the washing step, the peroxidase bound in the complex is developed by tetramethylbenzidine substrate. The protein concentration was determined by spectrophotometric measurements of the absorbance at 450 nm using a UV-Vis 1700 spectrophotometer (Shimadzu, Duisburg, Germany).
3
Chlorophyll Content in Leaf Tissue
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Leaves were harvested 50 days after transplanting for assessing chlorophyll content. One gram of leaf tissue was crushed in 80% pre-chilled acetone and the volume was made up to 100 mL. The absorbance of the supernatant was recorded at 645, 663, and 645 nm using a UV-vis 1700 spectrophotometer, Shimadzu, Japan. The amount of chlorophyll (mg/g) in leaf tissue was calculated by the formula given in Sadashivam and Manickam (1996) .
4
Spectroscopic Analysis of Cellulose-Based Films
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The light transmittance of the films was measured through a Shimadzu Corporation UV-2550 spectrophotometer. The study was conducted over a wavelength range of 200–600 nm. Small pieces (4.0 cm × 1.0 cm) were placed in the device cell, and air was used as a benchmark for transparency. UV–vis spectra of CG film, CG-ELE film, and CG-QUE were measured by Shimadzu UV–Vis 1700 Spectrophotometer. For the stated purpose, precise values of pH (pH 1–12) were achieved in eucalyptus extract solutions and films (1 × 1) by adjusting them with hydrochloric acid (HCl) or ammonia (NH3) 0.1 mol/L solutions. The film without ELE and QUE was employed as a blank.
5
Quantification of p53 Levels in Transfected U-87 Cells
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The cells were transfected following the procedure mentioned above. The medium was removed, and U-87 cells were washed (3 times) with phosphate buffer solution (PBS). Cells were then collected and the p53 content was quantified by using the p53 pan ELISA kit (Roche Applied Science, Penzberg, Germany). This kit is based on a sandwich enzyme-immunoassay. All of the experimental protocol steps, provided by the manufacturer, were followed. The levels of p53 were spectrophotometrically determined, at 450 nm, using a Shimadzu UV–vis 1700 spectrophotometer. Non-transfected cells and cells transfected with naked pDNA were considered as controls.
6
Proline Content Determination Protocol
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Proline content was measured by crushing 0.5 g sample in 10 mL 3% aqueous sulphosalicylic acid followed by filtering with Whatman no. 2 filter paper. In 2 mL of filterate, 2 mL glacial acetic acid and 2 mL acid ninhydrin was added and kept in a boiling water bath for a period of 1 h. The reaction was terminated by placing it in an ice bath. Further, toluene (4 mL) was mixed by stirring for 20–30 s, and the solution was kept at room temperature for toluene layer separation. The upper layer is taken, and the absorbance of the red color was taken at 520 nm with a UV–vis 1700 spectrophotometer, Shimadzu, Japan. Calculations were performed using a standard curve, as described in Sadashivam and Manickam [33 ].
7
Spectral Characterization of Carotenoids
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Absorption spectra of the carotenoids were measured by a UV–VIS 1700 spectrophotometer (Shimadzu) at room temperature. The characterization of the carotenoids was based on their absorption spectrum of the pigment extracts. The dried pigment extracts and pure carotenoids were diluted in 1 mL methanol and their spectra measured at wavelengths (λ) of 200–1100 nm. Identification of the pigments was based on their spectral properties and λ at maximum absorbance (λmax) as well as their comparative study to other references. FT-IR measurements of the purified carotenoids were performed with a Jasco FT/IR-6800 (Tokyo, Japan), by using the ATR (Diamond) method. The type of ATR used was the Jasco ATR Pro One. The pure carotenoids were diluted in acetone and dropped into the sample chamber and left until the solvent evaporated. Thereafter, 64 scans were accumulated with ranges from 400–4000 cm−1. The spectrum resolution was 4 cm−1. The ATR prism was ZnSe. The measurement was conducted at room temperature under dark conditions. Subsequently, the data obtained were analyzed using OriginPro 8.5.1 software (OriginLab, Northampton, MA, USA).
8
Quantification of p53 Protein Levels
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The p53 protein levels have been determined by using p53 pan ELISA kit (Roche Applied Science, Basel, Switzerland), following the procedure described by the manufacturer. The protein concentration was determined by spectrophotometrically measuring the absorbance at 450 nm using a UV-Vis 1700 spectrophotometer (Shimadzu, Tokyo, Japan). All the experiments were repeated three times in triplicate.
9
Antimicrobial Carrageenan-based Biopolymer Films
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All of the required chemicals, including methanol, ethanol, aluminum chloride, potassium acetate, ammonia, hydrochloric acid, κ-carrageenan, and quercetin, were purchased from Aldrich. The bacterial strains used in the research, i.e. S. aureus ATCC 6538, S. epidermidis ATCC 12228, B. subtilis ATCC 6633, E. coli ATCC 8739, P. aeruginosa ATCC 9027 and K. pneumonia ATCC 10031 procured from the Iranian Research Organization for Science and Technology (IROST). The κ-carrageenan/quercetin (CG-QUE) and carrageenan/ eucalyptus leaf extract (CG-ELE) films were analyzed using ZEISS Sigma 300 Field Emission Scanning Electron Microscope (FESEM), Shimadzu 8400 S Fourier Transform Infrared Spectrometer (FT-IR), and Shimadzu UV–Vis 1700 Spectrophotometer. The mechanical properties were tested using TA-XTPlus Texture Analyzer from Stable Micro Systems, Co., UK.
10
Chlorophyll Content Determination in Leaves
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After 45 days of transplantation, leaves were collected for assessing chlorophyll content as described by Witham et al. (1971) . One gram of leaf tissue was crushed in 80% pre-chilled acetone, and the volume was made up to 100 ml with pre-chilled acetone. The absorbance of the supernatant was recorded at 663 and 645 nm using UV–vis 1700 spectrophotometer (Shimadzu, Japan). The amount of chlorophyll present (mg/g) in the leaf tissue was calculated using the following formula:
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