Name: Solarbio Cell Iron Content Assay Kit 50T | 48S
Availability: InStock

Cell Iron Content Assay Kit

筆記: 有必要預測 2-3 正式測定前差異較大的樣品. 操作設備: 分光光度計

目錄號: BC5310

尺寸:50T/48S

成分:

萃取液: 液體 30 毫升×1. 2-8℃保存.

試劑一: 液體 15 毫升×1. 2-8℃保存. 試劑二: 液體 35 毫升×1. 2-8℃保存. 試劑三: 液體 6 毫升×1. 2-8℃保存.

標準: 液體 1 毫升×1. 2-8℃保存. 1 μmol/mL of Fe3+ standard solution. Add 200μL distilled water to 200μL Fe3+ standard solution of 1 使用前μmol/mL, and mix well to form the Fe3+ standard solution of 0.5 微摩爾/毫升.

產品描述:

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

需要但未提供的試劑和設備:

分光光度計, 桌上型離心機, 可調式移液器, 1毫升玻璃比色皿, cell ultrasonic crusher, 冰, 蒸餾水.

程式:

樣品製備:
將細菌或細胞收集到離心管中, and discard the supernatant after centrifugation. According to the proportion of bacteria or cell number (104): 萃取液體積 (毫升) 的 500-1000-1 提取. 建議 5 million bacteria or cells amount to 0.5mL of Extract solution. Split the bacteria or cell with ultrasonication (置於冰上, 超音波功率200W, 工作時間3秒, 間隔7秒, 重複 30 次). 離心機在 8000 克為 10 4℃分鐘去除不溶物, and take the supernatant on ice before
Ultrasonication working time could be prolonged properly if samples are fungi, 細菌, or other microorganisms with cell

二. 決心程式:

預熱分光光度計 30 分分鐘, 調整波長至 510 奈米, and set zero with distilled
Add reagents with the following:

試劑 (微升)	試管 (在)	空白管 (AB)	標準管 (作為)
樣本	100	–	–
蒸餾水	–	100	–
標準溶液 (0.5 微摩爾/毫升)	–	–	100
試劑一	200	200	200
試劑Ⅱ	600	600	600
試劑Ⅲ	100	100	100
充分混合, and place at 25℃ for 10 分分鐘. Take 1mL of reaction solution in 1mL glass cuvette. 測量吸光度 510 奈米, 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.

三、. Cell Iron Content 計算

Bacteria/cells number

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

2) 蛋白質專注

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, 0.5微摩爾/毫升;

VE: 萃取液體積, 0.5 毫升;

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

55.845: Relative molecular mass of Fe, 55.845ng/nmol; 500: 細菌或細胞總數, 5 百萬;

心肺復甦: 上清液樣品蛋白濃度, 毫克/毫升.

筆記:

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.
Sample supernatant protein concentration needs to be measured by yourself if cell iron content is calculated by protein

實驗範例:

拿 5 百萬個細胞, 添加 0.5 萃取液毫升數, 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

Solarbio Cell Iron Content Assay Kit 50T | 48S

描述

筆記: 有必要預測 2-3 正式測定前差異較大的樣品. 操作設備: 分光光度計