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Uv 1200 spectrophotometer

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The UV-1200 spectrophotometer is a laboratory instrument manufactured by Shimadzu. It is designed to measure the absorbance or transmittance of light through a sample over a range of ultraviolet and visible wavelengths.

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Lab products found in correlation

Atp bioluminescence assay kit, by Roche (3 mentions) Luminescence plate reader, by Molecular Devices (2 mentions) Ciras 3, by PP Systems (1 mentions)

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UV 1200 ultraviolet spectrophotometer UV-VIS 1200 spectrophotometer UV-1200 UV spectrophotometer

11 protocols using uv 1200 spectrophotometer

1

Phenolic Content Quantification Protocol

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The amount of total phenolics was determined by the procedure of Folin–Ciocalteu described by Amerine and Ough (1980) . The HE suspended in 0.5 mL 96% (v/v) ethanol was mixed with 30 mL of water and 2.5 mL of Folin–Ciocalteu’s reagent (Merck 9001, Darmstadt, Germany). After 30 s, 7.5 mL of 20% w/v sodium carbonate solution were added and the solution was mixed and diluted with water to a final volume of 50 mL. After 2 h in the dark at 20 °C, the absorbance of the samples was measured at 765 nm using a Shimadzu UV-1200 spectrophotometer (Shimadzu, Kyoto, Japan). The phenolic content was expressed in mg of gallic acid (GA) per gram of sample. The standard curve (50–750 mg L−1) was based on analytical grade GA (Sigma-Aldrich, Steinheim, Germany).
Fernández N.J., Damiani N., Podaza E.A., Martucci J.F., Fasce D., Quiroz F., Meretta P.E., Quintana S., Eguaras M.J, & Gende L.B. (2018). Laurus nobilis L. Extracts against Paenibacillus larvae: Antimicrobial activity, antioxidant capacity, hygienic behavior and colony strength. Saudi Journal of Biological Sciences, 26(5), 906-912.
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2

Preparation of RA-Incorporated GC Nanoparticles

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Preparation of the RA-incorporated GC nanoparticles was performed, as previously described by Jeong et al (10 (link)). A solution containing 5 mg RA in 1 ml DMF was slowly added to an aqueous solution containing 40 mg GC in 10 ml deionized water, whilst stirring. The stirring was continued for 20 min under darkened conditions. A dialysis membrane (molecular weight cut-off, 12,000 g/mol) was used to prepare the dialyzed solution in deionized water using a dialysis method for 1 day. The resulting dialyzed solution was lyophilized and analyzed. From the 20 ml solution, prepared by adding deionized water to the dialyzed solution, 100 µl was diluted with 9.9 ml DMSO. A UV-1200 spectrophotometer (Shimadzu Corporation, Kyoto, Japan) was then used to measure the drug contents at 365 nm, and empty GC vehicles were used as blank tests. The drug contents and loading efficiency were calculated using the following equations: Quantity of RA in the nanoparticles / weight of nanoparticles) × 100 and (residual quantity of RA in the nanoparticles / feeding quantity of RA) × 100, respectively.
LU H.C., MA J., ZHUANG Z., ZHANG Y., CHENG H.L, & SHI J.X. (2015). Retinoic acid-incorporated glycol chitosan nanoparticles inhibit the expression of Ezh2 in U118 and U138 human glioma cells. Molecular Medicine Reports, 12(5), 6642-6648.
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3

Spectrophotometric Analysis of Rice Amylose Content

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A simplified spectrophotometric method was applied to analyze the amylose content of rice.9) A rice sample was ground into a powder using a mortar and a pestle, and the powder (approximately 100 mg) was mixed with 1 mL of 95 % (v/v) ethanol and 9 mL of 1 mol/L sodium hydroxide in a test tube. The mixture was incubated for 10 min at 100 °C using an AHA hot plate (As One, Osaka, Japan), and then water was added to reach a final volume of 100 mL. Five mL of the solution was transferred into a clean test tube and was mixed with 1 mL of 1 mol/L acetic acid and 2 mL of iodine solution containing 0.2 % (w/v) iodine and 2 % (w/v) potassium iodide. Water was added to reach a final volume of 100 mL, and the absorbance of the solution was measured at 620 nm by using a UV-1200 spectrophotometer (Shimadzu Corporation, Kyoto, Japan). A calibration curve was prepared using potato amylose as a standard.
Sha Y, & Adachi S. (2016). Swelling Pressure of Wet Rice Grains Estimated from Distribution Coefficients of Saccharides. Journal of Applied Glycoscience, 63(2), 47-50.
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4

Colorimetric Determination of Plant Pigments

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Pigment content was measured colorimetrically according to Gratani17 . 0.5 g fresh levels dipped in 25 mL acetone alcohol mixture (acetone: alcohol = 1:1) until to turn white to extract chlorophyll a, b and carotenoid. The absorbance of extract liquor was determined by UV-1200 spectrophotometer (Shimadzu, Japan) at 645 nm (OD645), 663 nm (OD663) and 440 nm (OD440). The content of pigment was calculated as follow: chlorophyll a concentration (mg/L) = 12.7 × OD663 −2.69 × OD645; chlorophyll b concentration (mg/L) = 22.9 × OD645 −4.86 × OD663; total chlorophyll concentration (mg/L) = 8.02 × OD663 + 20.20 × OD645; carotenoid concentration (mg/L) = 4.7 × OD440 −0.27 × total chlorophyll concentration; chlorophyll a content (mg/g) = (chlorophyll a × 25 mL)/0.5 g; chlorophyll b content (mg/g) = (chlorophyll b × 25 mL)/0.5 g; carotenoid content (mg/g) = (carotenoid × 25 mL)/0.5 g.
Song J., Huang H., Hao Y., Song S., Zhang Y., Su W, & Liu H. (2020). Nutritional quality, mineral and antioxidant content in lettuce affected by interaction of light intensity and nutrient solution concentration. Scientific Reports, 10, 2796.
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5

Mitochondrial Enzyme Activity and ATP Assay

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Enzyme complex I (NADH-ubiquinone reductase), complex IV (cytochrome c oxidase, CcO) activity, and ATP levels were determined as described previously [7 (link)]. Briefly, cybrid cells were washed with ice-cold PBS, and then harvested, centrifuged, and suspended in 50 l of isolation buffer containing 250 mM sucrose, 20 mM HEPES, and 1 mM EDTA. Cell suspensions (containing ~3–4 mg of protein/ml) were added to a cuvette containing 0.95 ml of 1 × assay buffer (10 mM Tris-HCl, and 120 mM KCl), and the reaction volume was brought to 1.05 ml with the addition of 1× enzyme dilution buffer (10 mM Tris-HCl, pH 7.0). The reaction was then initiated by the addition of 50 μl of ferrocytochrome substrate solution (0.22 mM). The change in absorbance of cytochrome c at 550 nm was measured using a Shimadzu (Kyoto, Japan) UV1200 spectrophotometer. Activity is expressed as micromoles of cytochrome oxidized per min−1 mg−1 protein using an extinction coefficient of 18.64 mM−1 cm−1. ATP levels were determined using an ATP Bioluminescence Assay Kit (Roche) following the manufacturer’s instruction [9 (link), 25 (link)]. Briefly, cells were harvested, incubated on ice for 15 min, and centrifuged at 13,000 g for 10 min. ATP levels were measured using a Luminescence plate reader (Molecular Devices) with an integration time of 10 s.
Yu Q., Fang D., Swerdlow R.H., Yu H., Chen J.X, & Yan S.S. (2016). Antioxidants Rescue Mitochondrial Transport in Differentiated Alzheimer’s Disease Trans-Mitochondrial Cybrid Cells. Journal of Alzheimer's disease : JAD, 54(2), 679-690.
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6

Quantifying Mitochondrial Respiratory Function

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Activity of Cytochrome c oxidase, a key enzyme of respiratory chain complex IV, and ATP levels were determined as described previously [17 (link)]. Briefly, cybrid cells were washed with ice-cold PBS, and then harvested, centrifuged, and suspended in 50 μl of isolation buffer containing 250 mM sucrose, 20 mM HEPES, and 1 mM EDTA. Cell suspensions (containing 3–4 mg of protein/ml) were added to a cuvette containing 0.95 ml of 1 × assay buffer (10 mM Tris-HCl, pH 7.0 and 120 mM KCl), and the reaction volume was brought to 1.05 ml with the addition of 1 × enzyme dilution buffer (10 mM Tris-HCl, pH 7.0 and 250 mM sucrose). The reaction was then initiated by the addition of 50 μl of ferrocytochrome substrate solution (0.22 mM). The rate of change in absorbance at 550 nm was recorded immediately using a Shimadzu (Kyoto, Japan) UV1200 spectrophotometer programed for a 5 s delay and 10 s intervals for 6 readings. ATP levels were determined using an ATP Bioluminescence Assay Kit (Roche) following the manufacturer’s instruction as we previously described [8 (link)]. Briefly, cells were quickly harvested by ATP lysis buffer, incubated on ice for 30 min, and then centrifuged at 12,000 g for 10 min. ATP levels were measured by Luminescence plate reader (Molecular Devices). A 1.6 s delay time after substrate injection and 10 s integration time were used.
Du F., Yu Q, & Yan S.S. (2021). PINK1 Activation Attenuates Impaired Neuronal-Like Differentiation and Synaptogenesis and Mitochondrial Dysfunction in Alzheimer’s Disease Trans-Mitochondrial Cybrid Cells. Journal of Alzheimer's disease : JAD, 81(4), 1749-1761.
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7

Photosynthetic Pigment Quantification in Leaves

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The Chl content of the leaves was determined by 80% acetone extraction. A fresh 0.2 g sample of the third leaf blade was weighed and placed in a 20 mL test tube containing 5 mL of absolute ethanol and 5 mL of 80% acetone and incubated in darkness for 24 h. The optical density (OD) was measured using a UV-1200 spectrophotometer (Shimadzu, Japan) at 470 nm (OD470) for carotenoids, 663 nm (OD663) for Chl a, and 645 nm (OD645) for Chl b. These measurements were used to calculate the content of each respective pigment in the leaves using the following formulas [67 (link), 68 (link)]:
Chla(mg⋅g−1)=(12.72OD663nm−2.59OD645nm)V/1000W;Chlb(mg⋅g−1)=(22.88OD645nm−4.67OD663nm)V/1000W;Carotenoids(mg⋅g−1)=(1000OD470nm−3.27Chla−104Chlb)V/(229×1000W),
where V is the total volume of acetone extract (ml), and W is the FW (g) of the sample.
Gao S., Liu X., Liu Y., Cao B., Chen Z, & Xu K. (2020). Photosynthetic characteristics and chloroplast ultrastructure of welsh onion (Allium fistulosum L.) grown under different LED wavelengths. BMC Plant Biology, 20, 78.
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8

Mitochondrial Respiratory Chain Enzyme Activities

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The key enzyme activities associated with the mitochondrial respiratory chain, including complex I (NADH-ubiquinone reductase), complex II (succinate dehydrogenase), complex III (ubiquinol-cytochrome c reductase), complex IV (cytochrome c oxidase, CcO) and citrate synthase, were measured in cybrid cell lysates and isolated platelets’ mitochondria as described previously [31 –33 (link)]. ATP levels were measured using an ATP Bioluminescence Assay Kit (Roche) following the manufacturer’s instructions and using a Shimadzu (Kyoto, Japan) UV1200 spectrophotometer.
Gan X., Wu L., Huang S., Zhong C., Li G., Yu H., Swerdlow R.H., Chen J.X, & Yan S.S. (2014). Oxidative stress-mediated activation of extracellular signal-regulated kinase contributes to mild cognitive impairment-related mitochondrial dysfunction. Free radical biology & medicine, 75, 230-240.
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9

Chlorophyll and Carotenoid Extraction and Quantification

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The Chl content of the leaves was determined by 80 % acetone extraction. A fresh 0.2 g sample of the third leaf blade was weighed and placed in a 20 mL test tube containing 5 mL of absolute ethanol and 5 mL of 80 % acetone and incubated in darkness for 24 h. The optical density (OD) was measured using a UV-1200 spectrophotometer (Shimadzu, Japan) at 470 nm (OD 470 ) for carotenoids, 663 nm (OD 663 ) for Chl a, and 645 nm (OD 645 ) for Chl b.
These measurements were used to calculate the content of each respective pigment in the leaves using the following formulas [67, 68]:
Chl a (mg g -1 ) = (12.72 OD 663 nm -2.59 OD 645 nm ) V/1 000 W;
Chl b (mg g -1 ) = (22.88 OD 645 nm -4.67 OD 663 nm ) V/1 000 W;
Carotenoids (mg g -1 ) = (1 000 OD 470 nm -3.27 Chl a -104 Chl b) V/(229 × 1 000 W), where V is the total volume of acetone extract (ml), and W is the FW (g) of the sample.
Gao S., Liu X., Liu Y., Cao B., Chen Z., & Xu K. (2020). Photosynthetic characteristics and chloroplast ultrastructure of Welsh onion (Allium fistulosum L.) grown under different LED wavelengths.
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10

Chlorophyll and Carotenoid Quantification Protocol

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The Chl content of the leaves was determined by 80 % acetone extraction. A fresh 0.2 g sample of the third leaf blade was weighed and placed in a 20 mL test tube containing 5 mL of absolute ethanol and 5 mL of 80 % acetone and incubated in darkness for 24 h. The optical density (OD) was measured using a UV-1200 spectrophotometer (Shimadzu, Japan) at 470 nm (OD 470 ) for carotenoids, 663 nm (OD 663 ) for Chl a, and 645 nm (OD 645 ) for Chl b. These measurements were used to calculate the content of each respective pigment in the leaves using the following formulas [67, 68]:
Chl a (mg g -1 ) = (12.72 OD 663 nm -2.59 OD 645 nm ) V/1 000 W;
Chl b (mg g -1 ) = (22.88 OD 645 nm -4.67 OD 663 nm ) V/1 000 W;
Carotenoids (mg g -1 ) = (1 000 OD 470 nm -3.27 Chl a -104 Chl b) V/(229 × 1 000 W), where V is the total volume of acetone extract (ml), and W is the FW (g) of the sample.
Gao S., Liu X., Liu Y., Cao B., Chen Z., & Xu K. (2020). Photosynthetic characteristics and chloroplast ultrastructure of Welsh onion (Allium fistulosum L.) grown under different LED wavelengths.
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