Соларбио Малоновый диальдегид (МДА) Набор для анализа содержания

Малоновый диальдегид (МДА) Набор для анализа содержания

Примечание: It is necessary to predict 2-3 large difference samples before the formal determination. Операционное оборудование: Спектрофотометр.

Кот нет: BC0020

Размер: 50Т/48С

Компоненты:

Примечание: The working solution for MDA detection is difficult to dissolve, which can be heated at 70℃ and vibrated violently to promote dissolution. Or by ultrasonic treatment to promote dissolution.

Описание продукта:

• Lipid peroxide is generated through the interaction of oxygen free radicals with unsaturated fatty acids, eventually forming compounds such as malondialdehyde (МДА). The level of lipid peroxidation serves as an indicator, which can be assessed by measuring MDA levels.
• Under acidic and high-temperature conditions, a brown-red compound, 3,5,5-three methyl sulfamethoxazole-2,4-two ketone, is synthesized via a condensation reaction between MDA and thiobarbituric acid (TBA). This compound exhibits maximum absorption at 532 нм. The assessment of lipid peroxidation involves colorimetric analysis. Однако, the presence of soluble sugars can interfere with the detection process. The reaction of soluble sugars with TBA produces a color reaction with absorption wavelengths at 450 nm and 532 нм. In this assay kit, the MDA content is determined by the variance in absorbance at 532 нм, 450 нм, и 600 нм.
• Due to the presence of sucrose in plant tissues and glucose in animal tissues, the kit includes two computational formulas tailored for sucrose and glucose. These formulas are suitable for lipid assessment.

Требуемые реагенты и оборудование, которые не входят в комплект поставки:

Спектрофотометр, водяная баня, настольная центрифуга, пипетка для переноса,1 стеклянная кювета мл, ступка/гомогенизатор, лед и дистиллированная вода.

Процедура:

я. Базовые приготовления:

Бактерии или клетки:

Collect bacteria or cells into the centrifuge tube. 5 million bacteria or cells could be mixed with 1 mL of Extraction reagent. Use ultrasonication to split bacteria and cells (помещенный на лед, ultrasonic power 200W, ultrasonic time 3 секунды, интервал 10 секунды, повторить для 30 раз). Центрифуга в 8000 ×g для 10 minutes at 4℃ to remove insoluble materials, and take supernatant on ice before testing.

Салфетка:

0.1 g of tissue could be mixed with 1 mL of Extraction reagent and fully homogenized on ice bath. Then centrifuge at 8000 ×g для 10 minutes at 4℃ to remove insoluble materials, and take the supernatant on ice before testing.

сыворотка:

Detect

II. Определение процедура:

1. Preheat spectrophotometer for more than 30 минуты, set zero with distilled
2. Добавьте реагенты из следующего списка:

The mixture would be incubated at 100℃ for 60 минуты (tightly close to prevent moisture loss), cooled on ice, and centrifuged at 10000 ×g для 10 minutes at room temperature to remove insoluble materials. Take supernatant in 1 стеклянная кювета мл, and measure the absorbance at 450 нм, 532 nm and 600 нм.

∆A450=A450(Т)-A450(Б), ∆A532=A532(Т)-A532(Б), ∆A600 =A600(Т)-A600(Б). Blank tube needs to test once or twice.

III. Расчет:

1. Салфетка, bacteria or cultured cells

• Концентрация белка:

МДА (nmol/mg prot)"="(6.45×(∆A532-∆A600)-1.29×∆A450)×Врв÷(Cpr×Vs)

=5×(6.45×(∆A532-∆A600)-1.29×∆A450)÷Cpr

• Вес образца:

МДА (nmol/g weight)"="( 6.45×(∆A532-∆A600)- 1.29×∆A450)×Врв÷(W×Vs÷Vsv)

=5×(6.45×(∆A532-∆A600)- 1.29×∆A450)÷Вт

• Cellamount:

МДА (nmol/104cell)"="( 6.45 ×(∆A532-∆A600)- 1.29×A∆450)×Врв÷(400×Vs÷Vsv)

=0.01×(6.45×(∆A532-∆A600)- 1.29×∆A450)

• сыворотка:

МДА (nmol/mL)"="(6.45×(ΔA532 -ΔA600)-1.29×ΔA450)×Vrv÷Vs

=5×(6.45×(ΔA532 -ΔA600)-1.29×ΔA450)

2. Plants tissue:

• Вес образца

МДА (nmol/g weight)"=" (6.45×(∆A532-∆A600)-0.56×∆A450)×Врв÷(W×Vs÷Vsv)

=5×(6.45 ×(∆A532-∆A600)- 0.56×∆A450)÷Вт

• Концентрация белка:

МДА (nmol/mg prot)"="( 6.45 ×(∆A532-∆A600)- 0.56×∆A450)×Врв÷(Cpr×Vs)

=5×(6.45 ×(∆A532-∆A600)- 0.56×∆A450)÷Cpr

Веревка: Общий объем реакции, 1 мл; Против: Объем образца, 0.2 мл;

Всв: Объем экстракции, 1 мл;

КПР: Концентрация белка образца, мг/мл; Вт: Вес образца, г;

400: Total number of bacteria and cells, 5 миллион.

Примечание:

If it is found that the absorbance value of the sample is too low, the boiling water bath time can be adjusted from 60 минут до 90 minutes or longer. The detection of MDA in the same experiment needs to be extended to the same time to avoid errors.

Рекомендации

• Spitz D R, Oberley L W. An assay for superoxide dismutase activity in mammaliantissue homogenates[Дж]. Analytical Biochemistry,1989
• Masayasu M, Hiroshi Y. A simplified assay method of superoxide dismutase activity for clinical use[Дж]. Clinica Chimica

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