Solarbio Cell Iron Content Assay Kit 50T | 48S

Cell Iron Content Assay Kit

Bilješka: Potrebno je predvidjeti 2-3 Uzorci velike razlike prije formalnog određivanja. Operativna oprema: Spektrofotometar

Mačka ne: BC5310

Veličina:50T/48s

Komponente:

Otopina ekstrakta: Liquid 30 ml × 1. Store at 2-8℃.

Reagens i: Liquid 15 ml × 1. Store at 2-8℃. Reagens II: Liquid 35 ml × 1. Store at 2-8℃. Reagens iii: Liquid 6 ml × 1. Store at 2-8℃.

Standard: Liquid 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.

Opis proizvoda:

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 NM. According to the measure, absorbance at 520 nm can reflect cell iron concentration.

Potrebni reagensi i oprema, ali nisu pruženi:

Spektrofotometar, stolna centrifuga, Podesiva pipeta, 1ML staklena kiveta, cell ultrasonic crusher, led, destilirana voda.

Postupak:

1. Priprema uzorka:
2. Collect bacteria or cells into the centrifuge tube, and discard the supernatant after centrifugation. According to the proportion of bacteria or cell number (104): Extract solution volume (ml) od 500-1000-1 izdvojiti. Predlaže se da 5 million bacteria or cells amount to 0.5mL of Extract solution. Split the bacteria or cell with ultrasonication (stavljen na led, ultrasonic power 200W, Radno vrijeme 3s, Interval 7s, ponoviti 30 vrijeme). Centrifuga na 8000 g for 10 minuta u 4 ℃ za uklanjanje netopljivih materijala, 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. Odlučnost postupak:

1. Zagrijte spektrofotometar za 30 minuta, Podesite valnu duljinu na 510 NM, and set zero with distilled
2. Add reagents with the following:

Reagens (μL)	Epruveta (NA)	Prazna cijev (Ab)	Standard (KAO)
Uzorak	100	–	–
Destilirana voda	–	100	–
Standard solution (0.5 μmol/ml)	–	–	100
Reagens i	200	200	200
Reagens ⅱ	600	600	600
Reagens ⅲ	100	100	100
Temeljito promiješajte, and place at 25℃ for 10 minuta. Take 1mL of reaction solution in 1mL glass cuvette. Measure the absorbance at 510 NM, recorded as AT, Ab, and AS. ΔAT = AT-AB, ΔAS=AS-AB. Blank tubes and standard tubes only need to be tested once or twice.

Iii. Cell Iron Content Calculations

• Bacteria/cells number

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

2) Protein concentration

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: Extract solution volume, 0.5 ml;

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

55.845: Relative molecular mass of Fe, 55.845ng/nmol; 500: Total number of bacteria or cells, 5 milijun;

CPR: Supernatant sample protein concentration, mg/ml.

Bilješka:

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

Eksperimentalni primjer:

1. uzeti 5 milijun stanica, dodati 0.5 ML otopine ekstrakta, 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

Povezani proizvodi：

BC1730/BC1735 Serum Ferri Ion Content Assay Kit

BC2860/BC2865 Serum Total Iron Binding Capacity(TIBC) Komplet za analizu

BC4350/BC4355 Tissue Iron Content Assay Kit