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Total Iron Binding Capacity (TIBC) Colorimetric Assay Kit
SKU: E-BC-K071-S-50
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Total Iron Binding Capacity (TIBC) Colorimetric Assay Kit
| SKU # | E-BC-K071-S |
| Detection Instrument | Spectrophotometer (520 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | TIBC |
| Sample Type | Serum |
| Sensitivity | 0.03 mg/L |
| Detection Range | 0.03-50 mg/L |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 50 min |
| Precision | Average inter-assay CV: 4.700% | Average intra-assay CV: 3.400% |
| Other instruments required | Micropipettor, Water bath, Vortex mixer, Centrifuge |
| Other reagents required | Normal saline (0.9% NaCl) |
| Storage | 2-8℃ |
| Valid period | 12 months |
Images
R M Adel et al investigate the destructive effect of iron overload by silver nanocomposite. Total iron binding capacity (TIBC) in rat serum was determined using TIBC colorimetric assay kit (E-BC-K071-S).
Table. Effect of iron sucrose administration and subsequent co-treatments of DFX with Tr-CA and Ag@Tr-CA on serum Fe, TIBC, ferritin and Tf levels in different groups. Mean ± S.E.M. and % of change.
| Fe μmol/1 | % of change | TIBC μmol/1 | % of change | Ferritin ng/mL | % of change | Tf ng/mL | % of change | |
| Group 1 (control) | 19.10 ± 0.64 | ___ | 51.18 ± 0.27 | ___ | 4.50 ± 0.12 | ___ | 10.60 ± 0.09 | ___ |
| Group 2 (iron sucrose) | 56.23a ± 2.94 | 194.40% | 87.70a ± 1.60 | 71.36% | 6.833a ± 0.47 | 51.78% | 23.80a ± 1.97 | 124.53% |
| Group 3 (DFX) | 19.66b ± 0.31 | 2.93% | 60.40a,b ± 1.33 | 18.01% | 5.33b ±0.26 | 18.44% | 14.30a,b ± 0.40 | 34.91% |
| Group 4 (DFX + Tr-CA) | 21.23b ± 0.63 | 11.15% | 60.43a,b ± 2.52 | 18.07% | 4.70b ± 0.21 | 4.44% | 15.23a,b ± 0.13 | 43.68% |
| Group 5 (DFX + Ag@Tr-CA) | 20.37b ± 0.47 | 6.65% | 56.93a,b ± 2.42 | 11.23% | 4.73b ± 0.27 | 5.11% | 16.43a,b ± 0.92 | 55% |
By ANOVA, P < 0.05, LSD Test. a = P < 0.05 vs Group 1. b = P < 0.05 vs Group 2. a & b = The mean difference is significant at the 0.05 level.
TIBC was significantly changed after treated.
Dilution of Sample
It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (0.03-50 mg/L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 1 |
| Rat serum | 1 |
| Porcine serum | 1 |
| Rabbit serum | 1 |
| Chicken serum | 1 |
| Cynomolgus monkey serum | 1 |
Note: The diluent is double distilled water or normal saline (0.9% NaCl).
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).
Kit Components & Storage
| Item | Component | Size (50 Assays) | Storage |
| Reagent 1 | 100 mg/L Iron Standard Stock Solution | 7 mL ×1 vial | 2-8℃, 12 months |
| Reagent 2 | Chromogenic Agent A | Powder × 2 vials | 2-8℃, 12 month shading light |
| Reagent 3 | Chromogenic Agent B | Powder × 2 vials | 2-8℃, 12 month shading light |
| Reagent 4 | Chromogenic Agent C | 60 mL × 2 vials | 2-8℃, 12 months |
| Reagent 5 | Iron Absorbent | 50 mg × 50 vials | 2-8℃, 12 months |
Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.
Technical Data:
Parameter:
Intra-assay Precision
Three human serum samples were assayed in replicates of 20 to determine precision within an assay (CV = Coefficient of Variation).
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (mg/L) | 3.50 | 24.60 | 44.50 |
| %CV | 3.6 | 3.2 | 3.4 |
Inter-assay Precision
Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (mg/L) | 3.50 | 24.60 | 44.50 |
| %CV | 4.5 | 4.7 | 4.9 |
Recovery
Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 100%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (mg/L) | 13.8 | 34.7 | 45 |
| Observed Conc. (mg/L) | 13.7 | 34.0 | 46.4 |
| Recovery rate (%) | 99 | 98 | 103 |
Sensitivity
The analytical sensitivity of the assay is 0.03 mg/L. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.
Standard Curve
The OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:
| Concentration (mg/L) | 0 | 5 | 10 | 20 | 30 | 40 | 50 |
| Average OD | 0.001 | 0.130 | 0.253 | 0.515 | 0.755 | 1.034 | 1.283 |
| Absoluted OD | 0 | 0.129 | 0.252 | 0.514 | 0.754 | 1.033 | 1.282 |