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Biosystems İron Binding Reagent

Biosystems İron Binding Reagent

PRINCIPLE OF THE METHOD
Excess of Fe3+ is added to the sample to saturate serum transferrin.

Uncomplexed Fe3+ is precipitated with magnesium hydroxide carbonate and the iron bonded to protein in the supernatant is then spectrophotometrically measured1.

CONTENTS AND COMPOSITION
A. Reagent. 1 x 50 mL Iron chloride (III) 0.12 mmol/L.
B. Reagent. 3.10 g. Magnesium hydroxide carbonate (powder). To be dispensed using the enclosed plastic spoon.

STORAGE
Store at 2-30° C
Reagents are stable until the expiry date shown on the label when stored tightly closed and if contaminations are prevented during their use.
Indications of deterioration:
Reagent A: Presence of particulate material, turbidity.
Reagent B: Presence of moisture.

ADDITIONAL REAGENTS
These auxiliary reagents are to be used together with the iron reagents contained in the Biosystems Iron-ferrozine kit (cod 11509).

Biosystems İron Binding Reagent

ADDITIONAL EQUIPMENT
– Desktop centrifuge
– Analyzer, spectrophotometer or photometer able to read at 560 ± 20 nm.

SAMPLES
Serum or heparinized plasma collected by standard procedures. Do not use hemolyzed samples. The total iron binding capacity of serum or plasma is stable for 7 days at 2-8°C.

 

PROCEDURE
1. Pipette into labelled tubes (Note 1):
Sample 0.5 mL
Reagent (A) 1.0 mL
2. Mix thoroughly and let stand for 5-30 minutes at room temperature.
3. Add to each tube one spoonful of Reagent (B).
4. Mix thoroughly and let stand the tubes for 30-60 minutes at room temperature. During this time mix thoroughly several times.
5. Centrifuge at a minimum of 3000 r.p.m. for 10 minutes.
6. Carefully collect the supernatant (Note 2).
7. Measure the iron concentration in the supernatant, using the kit Iron-ferrozine (BioSystems, cod 11509).

CALCULATIONS
Total Iron Binding Capacity (TIBC)
TIBC = Iron concentration in the supernatant x 3 (dilution)
Iron saturation
100 x serum iron concentration
=iron saturation (%)

TIBC REFERENCE VALUES
Total Iron Binding Capacity (TIBC)2
Infant: 100-400 ^g/dL = 18-72 ^mol/L Adult: 250-425 ^g/dL = 45-76 ^mol/L
Iron saturation2
Men: 20-50%
Women: 15-50%

Brand : Biosystems

 

Biosystems İron Binding Reagent