Fish estrogen receptor ELISA kit operation method technical detection range
March 02, 2024
Fish estrogen receptor ELISA kit operation method technology detection range <br>operation steps
The required slats were taken out from the aluminum foil pouch after equilibrating for 20 min at room temperature, and the remaining slats were sealed back to 4 ° C with a ziplock bag.
Set standard and sample wells, standard wells with different concentrations of standard 50μL;
Add 50 μL of the sample to be tested to the sample well; blank holes are not added.
In addition to the blank wells, 100 μL of horseradish peroxidase (HRP)-labeled detection antibody was added to each well of the standard well and the sample well, and the reaction well was sealed with a sealing plate, and incubated at 37 ° C in a water bath or incubator for 60 min.
Discard the liquid, pat dry on the absorbent paper, fill each well with the washing solution (350 μL), let stand for 1 min, remove the washing solution, pat dry on the absorbent paper, and repeat the washing 5 times (can also be washed with a washing machine) .
50 μL of each of the substrates A and B was added to each well, and incubated at 37 ° C for 15 min in the dark.
50 μL of the stop solution was added to each well, and the OD value of each well was measured at a wavelength of 450 nm within 15 min.
Fish estrogen receptor ELISA kit method of detection technology detection range kit performance
Detection range: 12.5 pg/mL – 400 pg/mL.
Sensitivity: The minimum detection concentration is less than 1.0 pg/mL.
Specificity: Does not cross-react with other soluble structural analogs.
Repeatability: The intra-plate variation coefficient is less than 10%, and the inter-plate variation coefficient is less than 15%.
Kit composition
name
96-well configuration
48-hole configuration
Remarks
Microporous plate
8 holes × 12
8 holes × 6
no
Standard
0.3mL*6 tube
0.3mL*6 tube
no
Sample diluent
6mL
3mL
no
Detection antibody-HRP
10mL
5mL
no
20× washing buffer
25mL
15mL
Dilute according to the instructions
Substrate A
6mL
3mL
no
Substrate B
6mL
3mL
no
Stop solution
6mL
3mL
no
Sealing film
2 sheets
2 sheets
no
Instruction manual
1 serving
1 serving
no
Ziplock bag
1
1
no
Remarks: 1. Standard concentration is: 400, 200, 100, 50, 25, 12.5 pg/mL 2. After a large number of normal specimen tests, the normal concentration values ​​of the specimens are within the detection range provided by the kit, and 50 μL of the sample can be directly loaded during the experiment. When part of the sample value exceeds the maximum standard concentration, the sample may be diluted with the sample dilution before the experiment.
Precautions
Incubation is carried out strictly in accordance with the specified time and temperature to ensure accurate results. All reagents must reach room temperature 20-25 °C before use. Store the reagents refrigerated immediately after use.
Incorrect cleaning can result in inaccurate results. Make sure to drain the liquid in the well as much as possible before adding the substrate. Do not let the micropores dry during the incubation.
Eliminate residual liquid and fingerprints at the bottom of the board, otherwise it will affect the OD value.
The substrate coloring solution should be colorless or very light, and the substrate liquid that has turned blue cannot be used.
Avoid cross-contamination of reagents and specimens to avoid erroneous results.
Avoid direct exposure to strong light during storage and incubation.
After balancing to room temperature, the sealed bag was opened to allow water-repellent drops to coagulate on the cold strip.
Any reagents should not be exposed to strong gases emitted by bleaching solvents or bleaching solvents. Any bleaching component will destroy the biological activity of the reagents in the kit.
Expired products cannot be used.
If the disease is likely to spread, all samples should be managed and the sample and test device processed in accordance with the prescribed procedures.
