- Catalogue No.
K201
- Size
100T(96 samples)
- Kit component
|
Item
|
Quantity
|
Instructions
|
Storage
|
|
Standard
|
Liquid 2mL×1 bottle
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1000 μmol/L Fe³+ standard, diluted 8 times to 125μmol/L standard before use.
|
2-8℃ for 3 months
|
|
Reagent 1
|
Powder×2 bottles
|
Add 7.5mL distilled water to dissolve fully before use.
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2-8℃ for 3 months
|
|
Reagent 2
|
Powder×2 bottles
|
Add 235μL glacial acetic acid and 7.5 mL distilled water to dissolve fully before use.
|
2-8℃ for 3 months
|
- Principle of the Assay
Serum iron refers to the iron bound by blood transferrin, which is often used to distinguish iron deficiency from non-iron deficiency anemia. Serum Fe3+ is reduced by sodium sulfite to form Fe2+, and Fe2+ is further colored with 2, 2 '-bipyridine. There is an absorption peak at 520nm, and the serum iron content could be calculated by measuring the light absorption value at this wavelength.
- Detection method
Micromethod
- Materials Not Supplied
Centrifuge, Adjustable pipette, Visible spectrophotometer/Microplate reader, Microglass cuvettes/96-well plate, Glacial acetic acid, Chloroform, Distilled water.
- Assay Procedure (For reference)
1.Sample preparation
Serum (plasma): Directly detect.
2.Add each reagent in turn according to the operation table
Before the formal test, 2-3 samples with significant differences should be selected for pre-test.
(1)Preheat visible spectrophotometer or microplate reader for more than 30 min, adjust the wavelength to 520 nm and the distilled water to zero.
(2)Add each reagent in turn according to the operation table
|
Reagent（µL）
|
Sample tube
|
Standard tube
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Blank tube
|
|
Serum /plasma
|
125
|
-
|
-
|
|
125μmol/mL standard
|
-
|
125
|
-
|
|
Distilled water
|
-
|
-
|
125
|
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Reagent 1
|
125
|
125
|
125
|
|
Reagent 2
|
125
|
125
|
125
|
|
Mix well, cover tightly, place in boiling water bath for 5min, cool with tap water. Add 62 μL chloroform（self-prepared）and mix thoroughly. At room temperature 10000rpm, centrifuge for 10min, carefully absorb 210 μL of the upper liquid, add it into the microglass cuvettes/96-well plate, and immediately measure the absorbance at 520 nm, recorded as A blank, A sample, and A standard.
|
3.Calculation
The content of serum iron(μmol /L)=[C standard×(A sample- A blank) ÷ (A standard- A blank)]
=125×(A sample- A blank) ÷(A standard- A blank)
C standard: 125 μmol/L Fe3+ standard
- Notes
1.The content of serum iron is low, and the vessels used (EP tubes) need to be paid attention to avoid being contaminated by iron.
2.If the absorbance value of the sample is greater than 0.5, it is recommended to dilute the sample with distilled water before measurement.
3.Sensitivity: 0.99 μmol/L
4.Range: 3.9-250 μmol/L
