Physical properties
Each liquid component is clear and transparent, without precipitation or floccules. Microplate aluminum foil bagIt should be vacuum packed without damage or leakage.
Use
For scientific research use only, for qualitative detection of human Epstein-Barr virus early antigen IgM antibodies (EA-IgM) in serum, plasma, and cell culture supernatant.
Specificity
This kit recognizes natural and recombinant human EB virus early antigen IgM antibodies (EA-IgM) and has no cross-reactivity with structural analogs.
Repeatability
The coefficient of variation within and between plates was<10%.
Test equipment required for the test
1, standard specification ELISA reader. 2automatic plate washer. 3, oscillator. 4, series of adjustable pipettes and tips. When testing a large number of samples at one time, it is best to use a multi-channel pipette.
Running program
1, all reagents and components should be restored to room temperature first, and it is recommended to do duplicate wells for positive and negative controls and samples. 22. Prepare the working solutions of various components of the kit according to the method described in the previous reagent preparation. 3, take out the required strips from the aluminum foil bag, seal the remaining strips in a ziplock bag and put them back in the refrigerator. 4, set up positive and negative controls, sample wells and blank wells, add positive and negative controls to each of the positive and negative control wells50μL, add the appropriately diluted sample to the sample well50μL, do not add any blank wells, and cover the reaction plate with a sealing film.37℃ water bath or incubator30min. 5remove the sealing film, discard the liquid, pat dry on absorbent paper, fill each well with washing solution, and let it stand20s, shake off the detergent and pat dry on absorbent paper. Repeat this process.4times (total wash plate5If using an automatic plate washer, follow the plate washer operating procedures to wash the plate and add soaking20sAfter washing the plate, pat the plate dry on a clean, lint-free paper before adding the substrate. 6, In addition to the blank wells, the positive and negative control wells and sample wells, add horseradish peroxidase-labeled detection antibodies100μL. 7, Cover the reaction plate with sealing film,37Incubate in a water bath or incubator away from light30min. 8, remove the sealing film, discard the liquid, pat dry on absorbent paper, fill each well with washing solution, and let it stand20s, shake off the detergent and pat dry on absorbent paper. Repeat this process.4times (total wash plate5If using an automatic plate washer, follow the plate washer operating procedures to wash the plate and add soaking20sAfter washing the plate, pat the plate dry on a clean, lint-free paper before adding the substrate. 9, the substrateAandBaccording to1:1Mix the volume thoroughly and add the substrate mixture to all wells.100μL. Cover the reaction plate with a sealing film.37Incubate in a water bath or incubator away from light15min. 10, add stop solution to all wells50μL,exist450nmRead the absorbance of each well on an ELISA reader (ODvalue).
Announcements
1)This kit is for in vitro research use only and is not intended for clinical diagnosis. 2)Please wear lab coats and latex gloves for protection during the test. Especially when testing blood or other body fluid samples, please follow the national biological laboratory safety protection regulations. 3)Strictly follow the specified time and temperature to incubate to ensure accurate results. All reagents must reach room temperature before use. 20-25℃. Refrigerate the reagent immediately after use. 4)Improper plate washing can lead to inaccurate results. Be sure to aspirate as much liquid as possible before adding substrate. Do not allow the wells to dry out during incubation. 5)Eliminate residual liquid and fingerprints on the bottom of the board, otherwise it will affect ODvalue. 6)The substrate developer solution should be colorless or very light in color. 7)Avoid cross contamination of reagents and specimens to avoid erroneous results. 8)Avoid direct sunlight during storage and incubation. 9)The sealed bag was opened after equilibration to room temperature to prevent water droplets from condensing on the cold strips. 10)Any reagent should not be exposed to bleaching solvents or the strong gases emitted by bleaching solvents. Any bleaching components will destroy the biological activity of the reagents in the kit. 11)The ELISA reader used for the test needs to be installed with 450±10nmWavelength filters with optical density range of 0-3.5It is recommended to use in advance 15Minutes to preheat. 12)Do not mix or substitute reagents in this kit with reagents from other lots or other sources. 13)The test used EPBoth the tube and the pipette tip are for single use and mixing is strictly prohibited. 14)Do not use expired reagents.
Problem analysis
①Problem description: The positive and negative control results are unstablePossible causes and corresponding countermeasures: 1.Inaccurate aspiration or addition of liquid——Check the pipette and tips 2.Equilibration time is too short——Ensure adequate balancing time 3.Incomplete washing——Ensure the washing time, washing times and liquid volume per well ②Problem description: The color is very weak or colorlessPossible causes and corresponding countermeasures: 1.Incubation time is too short——Ensure adequate incubation time 2.Incorrect test temperature——Use the recommended test temperature 3.Insufficient reagent volume or missed addition/Incorrect dilution——Check the aspiration and addition process to ensure that all reagents are added in order and in sufficient quantities. 4.Enzyme marker inactivation or substrate failure——Mix the enzyme conjugate and substrate and check and judge by rapid color development ③Problem description: Low readingsPossible causes and corresponding countermeasures: 1.The microplate reader is not set up correctly——Check wavelength and filter settings on the microplate reader/Turn on the ELISA reader in advance to preheat ④Problem description: Large coefficient of variationPossible causes and corresponding countermeasures: 1.Incorrect addition of liquid——Check the filling status ⑤Problem description: High background valuePossible causes and corresponding countermeasures: 1.The working concentration of the detection antibody is too high——Use the recommended dilution factor 2.Incomplete washing of ELISA plate——Ensure that each step is cleaned completely; if using an automatic plate washer, check whether all outlets are blocked; whether the washing solution provided in the kit is used 3.The washing liquid is contaminated——Prepare new lotion ⑥Problem description: Low sensitivityPossible causes and corresponding countermeasures: 1.ELISAImproper storage of the test kit——Store relevant reagents according to the instructions 2.Not terminated before reading——ODStop solution should be added to each well before reading.
Statement
The kit is for research use only. If it is used for clinical diagnosis or any other purpose, our company will not be responsible for any problems arising therefrom and will not bear any legal liability.
