Solarbio Cell Iron Content Assay Kit 50T | 48S

Cell Iron Content Assay Kit

Nota: Es necesario predecir 2-3 muestras de gran diferencia antes de la determinación formal. Equipo de operación: espectrofotómetro

No gato: BC5310

Tamaño:50T/48S

Componentes:

Extraer solución: Líquido 30 mL×1. Store at 2-8℃.

reactivo yo: Líquido 15 mL×1. Store at 2-8℃. Reactivo II: Líquido 35 mL×1. Store at 2-8℃. Reactivo III: Líquido 6 mL×1. Store at 2-8℃.

Estándar: Líquido 1 mL×1. Store at 2-8℃. 1 μmol/mL of Fe3+ standard solution. Add 200μL distilled water to 200μL Fe3+ standard solution of 1 μmol/mL before use, and mix well to form the Fe3+ standard solution of 0.5 µmol/mL.

Descripción del Producto:

Iron is one of the essential trace elements in the human body, which is the main component of hemoglobin, myoglobin, cytochrome, and other enzyme systems. Iron can assist in the transport of oxygen and promote fat oxidation. Iron deficiency can easily cause anemia, and metabolic disorders, and affect the immune function of the body.

Fe3+ is reduced by hydroxylamine hydrochloride to Fe2+. Fe2+ could react with Phenanthroline Monohydrate to form a kind of orange compound under weak acid conditions, which has an absorption peak at 510 Nuevo Méjico. According to the measure, absorbance at 520 nm can reflect cell iron concentration.

Reactivos y equipos necesarios pero no suministrados:

espectrofotómetro, centrífuga de escritorio, pipeta ajustable, 1cubeta de vidrio de ml, cell ultrasonic crusher, hielo, distilled water.

Procedimiento:

1. preparación de la muestra:
2. Recoja bacterias o células en el tubo de centrífuga., and discard the supernatant after centrifugation. According to the proportion of bacteria or cell number (104): Volumen de solución de extracción (mililitros) de 500-1000-1 to extract. It is suggested that 5 million bacteria or cells amount to 0.5mL of Extract solution. Split the bacteria or cell with ultrasonication (colocado sobre hielo, potencia ultrasónica 200W, working time 3s, intervalo 7s, repetir 30 veces). Centrifugar en 8000 g for 10 minutos a 4 ℃ para eliminar materiales insolubles, and take the supernatant on ice before
3. Ultrasonication working time could be prolonged properly if samples are fungi, bacteria, or other microorganisms with cell

II. Determinación procedimiento:

1. Precalentar el espectrofotómetro durante 30 minutos, ajustar la longitud de onda a 510 Nuevo Méjico, and set zero with distilled
2. Add reagents with the following:

III. Cell Iron Content Cálculos

• Bacteria/cells number

Cell iron content (ng/104 cell) =(Cs×ΔAT÷ΔAS)×VE×103×55.845÷500=27.922×ΔAT÷ΔAS

2) Proteína concentración

Cell iron content (ng/mg prot)

=(Cs×ΔAT÷ΔAS)×VE×103×55.845÷(Cpr×VE)=27922.5×ΔAT÷ΔAS÷Cpr

Cs: Concentration of Fe3+ standard solution, 0.5µmol/mL;

VE: Volumen de solución de extracción, 0.5 mililitros;

103: Unit conversion factor, 1 μmol=103 nmol;

55.845: Relative molecular mass of Fe, 55.845ng/nmol; 500: Número total de bacterias o células., 5 millón;

RCP: Supernatant sample protein concentration, mg/mL.

Nota:

1. If ΔAT＜01, it is recommended to increase the sample supernatant size before determination. If AT＞1, it is recommended to dilute the sample supernatant with distilled water before determination. And modify the calculation formula.
2. Sample supernatant protein concentration needs to be measured by yourself if cell iron content is calculated by protein

Ejemplo experimental:

1. Llevar 5 millones de células, agregar 0.5 mL de solución de extracto, and split with ultrasonication. Centrifuge and take the supernatant. Then operate according to the determination steps, calculate ΔAT=AT-AB= 0.171- 0.000=0.171, ΔAS=AS-AB=0.573-0.000=0.573. The result is calculated according to cell numbers: Cell iron content (ng/104 cells) =27.922×ΔAT÷ΔAS=8.333ng/104 cell

Productos relacionados：

BC1730/BC1735 Serum Ferri Ion Content Assay Kit

BC2860/BC2865 Serum Total Iron Binding Capacity(TIBC) Kit de ensayo

BC4350/BC4355 Tissue Iron Content Assay Kit