Solarbio Cell Iron Content Assay Kit 50T | 48S

Cell Iron Content Assay Kit

Catatan: Hal ini perlu untuk diprediksi 2-3 perbedaan sampel yang besar sebelum penentuan formal. Peralatan Operasi: Spektrofotometer

Kucing No: BC5310

Ukuran:50T/48S

Komponen:

Ekstrak larutan: Cairan 30 mL×1. Store at 2-8℃.

Reagen I: Cairan 15 mL×1. Store at 2-8℃. Reagen II: Cairan 35 mL×1. Store at 2-8℃. Reagen III: Cairan 6 mL×1. Store at 2-8℃.

Standar: Cairan 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 mol/mL.

Deskripsi Produk:

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.

Reagen dan Peralatan Diperlukan tetapi Tidak Disediakan:

Spektrofotometer, mesin sentrifugal meja, pipet yang dapat disesuaikan, 1mL kuvet kaca, cell ultrasonic crusher, Es, air suling.

Prosedur:

1. Persiapan sampel:
2. Kumpulkan bakteri atau sel ke dalam tabung centrifuge, and discard the supernatant after centrifugation. According to the proportion of bacteria or cell number (104): Ekstrak volume solusi (ml) dari 500-1000-1 untuk mengekstrak. Disarankan demikian 5 million bacteria or cells amount to 0.5mL of Extract solution. Split the bacteria or cell with ultrasonication (ditempatkan di atas es, daya ultrasonik 200W, waktu kerja 3 detik, interval 7 detik, mengulang 30 waktu). Sentrifugasi di 8000 g untuk 10 menit pada suhu 4℃ untuk menghilangkan bahan yang tidak larut, and take the supernatant on ice before
3. Ultrasonication working time could be prolonged properly if samples are fungi, bakteri, or other microorganisms with cell

II. Tekad prosedur:

1. Panaskan terlebih dahulu spektrofotometer untuk 30 menit, sesuaikan panjang gelombangnya 510 nm, and set zero with distilled
2. Add reagents with the following:

AKU AKU AKU. Cell Iron Content Perhitungan

• Bacteria/cells number

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

2) Protein konsentrasi

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, 0.5mol/mL;

VE: Ekstrak volume solusi, 0.5 ml;

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

55.845: Relative molecular mass of Fe, 55.845ng/nmol; 500: Jumlah total bakteri atau sel, 5 juta;

Cpr: Supernatant sample protein concentration, mg/mL.

Catatan:

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

Contoh eksperimental:

1. Mengambil 5 juta sel, menambahkan 0.5 ML larutan ekstrak, 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

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