Zinc is one of the essential trace elements and plays an important role in insulin and porphyrin metabolism. This kit is suitable for detecting zinc levels in serum, plasma, urine, and milk.
The detection principle of this kit is as follows: under specific pH conditions, zinc ions form a stable, colorimetric complex with 5-Br-PADAP, the intensity of which is directly proportional to the concentration of zinc ions.
Kit Contents
Z1509014 | Components | Appearance | 100T | Storage | Quantity Per Test |
Z1509014A | Protein Precipitator | Liquid | 10 mL | 2-8℃. Store in the dark. | As needed |
Z1509014B | Chromogenic Agent | Liquid | 100 μL | 2-8℃. Store in the dark. | 1 μL |
Z1509014C | Buffer Solution | Liquid | 20 mL | 2-8℃ | 200μL |
Z1509014D | Standard Solution (1.54 mmol/L) | Liquid | 1 mL | 2-8℃. Store in the dark. | As needed |
Instructions for Use
1. Preparation of Sample Supernatant
Mix the sample with protein precipitation reagent in a 1:1 ratio, centrifuge at 4°C and 13,780 × g for 10 minutes, then collect the supernatant for testing.
Note: The supernatant after centrifugation must be clear. If turbidity is present, transfer the turbid supernatant to a new centrifuge tube and centrifuge again.
2. Preparation of standards
Using deionized water, dilute the standards (1.54 mmol/L) to 0, 1.54, 7.7, 15.4, 23.1, 30.8, 38.5, and 46.2 μmol/L according to the table below.
Serial Number | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
Concentration ( μmol/L) | 0 | 1.54 | 7.7 | 15.4 | 23.1 | 30.8 | 38.5 | 46.2 |
Standard solution (μL) | 0 | 1 | 5 | 10 | 15 | 20 | 25 | 30 |
Deionized water (μL) | 1000 | 999 | 995 | 990 | 985 | 980 | 975 | 970 |
3. Preparation of Chromogenic Working Solution
Mix Chromogenic Agent and Buffer Solution at a volume ratio of 1:200. Prepare immediately before use.
4. Sample Measurement
Carry out the sample measurement according to the table below (with the reaction taking place in a 96-well plate).
Reagent name | Standard well | Sample well |
Sample (μL) | - | 50 |
Standard solutions of different concentrations (μL) | 50 | - |
Chromogenic Working Solution (μL) | 200 | 200 |
Mix well, incubate for 5 min, then measure the OD value at 560 nm using a microplate reader. |
5. Result calculation
Standard curve equation: y = ax + b
Zinc Content Calculation: Zinc content (μmol/L) = (ΔA - b) / a × 2* × f
y: measured OD value minus blank OD value (OD at zero standard concentration);
x: standard concentration;
a: slope of the standard curve;
b: intercept of the standard curve;
ΔA: Sample OD value - blank OD value (OD value at standard concentration = 0);
2*: Dilution factor during sample pretreatment;
f: dilution factor applied to the sample prior to addition into the assay system.
Precautions
1. The sample must not cause hemolysis; metal chelating agents such as EDTA and citrate cannot be used as anticoagulants.
2. The used containers and reagents should be free from zinc ion contamination.
3. Before the formal test, it is recommended to select 2-3 samples with significant expected differences for a preliminary experiment.
4. For your safety and health, please wear a lab coat and latex gloves during the operation.
