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铁检测试剂盒
Iron Assay
品牌:
ScienCell
货号:
8448
价格:
$305
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Iron is an element that plays an essential role in a number of biological processes. Iron is bound to several proteins such as the oxygen-transporting hemoglobin in the red blood cells and oxygen storage protein myoglobin in the muscle. It is also stored by ferritin and hemosiderin. Inorganic iron contributes to redox reactions in the iron-sulfur clusters of many enzymes, such as nitrogenase. In this colorimetric assay, ferric carrier protein will dissociate ferric into solution under acidic conditions. After they are reduced to the ferrous ions, these ions react with chromogen to produce a stable colored complex and give absorbance at 590 nm. The color intensity thus formed is directly proportional to the concentration of iron in the sample. A specific chelate chemical is included in the buffer to block copper ion interference.
货号8448
产地美国
缩写IR
规格 100 tests
用途科研
储存-20度
运输干冰
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Question:

Hello, I wanted some clarification for part B: preparation of test samples for the Iron Assay Kit (Catalog #8448). I am confused what it means by " 4 volumes of the assay buffer", and "93 volumes of assay buffer (8448a), 2 volumes reductant (8448e) and 5 volumes sample reagents (8448d)." Does volumes just mean any unit of volume as long as that ratio is used? Thank you, Lauren Nardacci

Answers:

  • Posted by Manoj Sharma on Wednesday, January 6, 2016

    Hi Lauren,

    We have revised the preparation as follows:

    B. Preparation of test samples
    1. One volume of cells or tissues can be homogenized in 4 volumes of the assay buffer (8448a). Centrifuge the samples at 13,000 ×g for 10 minutes to remove insoluble material.
    2. Prepare a fresh working buffer by mixing assay buffer (8448a), reductant (8448e) and sample reagents (8448d) at 93:2:5 ratio (volume ratio). (e.g., 930 μL mixing assay buffer, 20 μL reductant, and 50 μL sample reagents)
    3. Mix sample (cell lysate or serum) with equal volume of working buffer.
    4. Centrifuge the mixed sample 13,000 ×g for 5 minutes and take the supernatant as the test samples for the iron assay.
    5. Test samples should be serial diluted to make sure the readings are within the standard curve range. Prepare test samples to a final volume of 50 μL/well on the 96-well flat bottom plate.

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