Application

This kit can be used to measure the Total Iron Binding Capacity (TIBC) content in serum samples. 

Detection significance

Total iron binding capacity (TIBC) was used as a parameter to evaluate the maximum capacity of serum iron transport. Iron is an essential biological element in organisms because it is involved in many metabolic processes such as oxygen transport, DNA synthesis and electronic transport. TIBC is also indirectly used to assess the level of serum transferrin.

Detection principle

The excess iron is added to the serum to bind all the ferritin in the serum, and the excess iron is adsorbed by adding the iron adsorbent. The iron bind with the ferritin is separated from the protein by the action of acid solution and reductant. Fe3+ in serum is reduced to Fe2+, Fe2+ binds with bipyridine to form pink complex. In a certain range, the amount of TIBC is positively correlated with the depth of color. The iron content measured is, minus serum iron value, which is called unsaturated iron binding force. Total iron binding capacity minus serum iron value is unsaturated iron binding capacity (UIBC).

Experimental instruments

Test tube, Micropipettor, Vortex mixer, Centrifuge, Water bath, Spectrophotometer (520 nm)

The preparation of sample

1. Collect the serum sample by conventional method. It is recommended to take 2~3 samples which expected large difference to do pre-experiment before formal experiment.

2. Take 1 mL of serum, add 1 mL of 10 mg/L Iron Standard application solution, mix fully and stand at room temperature for 10 min. Then add a vial of reagent 5, mix fully and stand at room temperature for 5 min, repeat the mix and stand steps for 4 times. Centrifuge at 2300 g for 10 min and take the supernatant for detection.

Operation steps

1. Blank tube: Add 1.0 mL of Double-distilled water into a 5 mL EP tube.

Standard tube: Add 1.0 mL of 1 mg/L Iron Standard application solution into a 5 mL EP tube.

Sample tube: Add 1.0 mL of sample into a 5 mL EP tube.

2. Add 2.0 mL of Chromogenic agent into each tube. Oscillate fully and incubate in 100℃ water bath for 5 min.

3. Cool the tubes with running water, then centrifuge at 2300 g for 10 min (If the supernatant is turbid, collect the turbid supernatant into another new EP tube and centrifuge again). Take 1.0 mL of the supernatant.

4. Set spectrophotometer to zero with double-distilled water and measure the OD value of each tube at 520 nm wavelength with 0.5 cm optical path cuvette.

Note: It can be refer to the following operating table.

Technical parameters

1. The sensitivity of the kit is 0.03 mg/L.

2. The intra-assay CV is 3.4 % and the inter-assay CV is 4.7 %.

3. The recovery of the kit is 100 %.

4. The linear range of the kit is 0.03-50 mg/L.

Notes

1. This kit is for research use only.

2. Instructions should be followed strictly, changes of operation may result in unreliable results.

3. The valid of kit is 6 months.

4. Do not use components from different batches of kit.