Instruction Manual for Influenza A IgM (FLU-A IgM) ELISA Kit
Instruction Manual for Influenza A IgM (FLU-A IgM) ELISA Kit Generic name: Influenza A IgM (FLU-A IgM) ELISA kit purpose of usage: This kit qualitatively determines influenza A type IgM (FLU-A IgM) in blood or other related tissues Experimental principle: This kit uses indirect enzyme-linked immunoassay (ELISA) to determine influenza A type IgM (FLU-A IgM) in the specimen. use The purified influenza A IgM (FLU-A IgM) antibody is coated on a microplate to make a solid-phase antibody that can be combined with influenza A in the sample. Type IgM (FLU-A IgM) combined, washed to remove unbound influenza type IgM (FLU-A IgM) and other components After separation, it is combined with HRP-labeled goat anti-human antibody to form an antibody-antigen-enzyme-labeled antibody complex, which is washed thoroughly Add substrate TMB to develop color. TMB is converted into blue under the catalysis of HRP enzyme, and into the final yellow under the action of acid color. Measure the absorbance (OD value) at 450nm with a microplate reader and compare it with the CUTOFF value to determine the specimen The presence or absence of influenza A type IgM (FLU-A IgM). Kit composition: 1. Specimen processing: (1) Serum and plasma specimens: can be directly detected 2) Other specimens: prepared according to relevant literature. ( 2. Perform the experiment as soon as possible after the specimen is prepared as required. If it can not be detected in time, the specimen can be stored at -20 ℃ for one month, However, repeated freezing and thawing should be avoided. 3. The sample containing NaN3 cannot be detected because NaN3 inhibits horseradish peroxidase (HRP) activity. Steps: 1. Numbering: The microwells corresponding to the samples are numbered in order. Each plate should be set with 2 wells for negative control, 2 wells for positive control, and 1 blank control. Wells (the blank control wells do not add samples and enzyme reagents, the remaining steps are the same) 2. Add sample: add 50μl of negative control and positive control (standard) to the negative and positive control wells respectively. Then under test Add 40μl of sample diluent to the sample well, and then add 10μl of the sample to be tested. Add sample to the bottom of the well of the microplate, Try not to touch the wall of the hole, shake gently to mix, 3. Incubation: seal the plate with the sealing film and incubate at 37 ° C for 30 minutes. 4. Mixing solution: add 20 times concentrated washing liquid to distilled water to 600ml and reserve 5. Washing: Carefully peel off the sealing film, discard the liquid, spin dry, fill each well with washing liquid, let stand for 30 seconds and then discard, so Repeat 5 times and pat dry. 6. Add enzyme: add 50μl of enzyme label reagent (or one drop) to each well, except for blank wells. 8. Washing: The operation is the same as 5. 9. Color development: add 50μl (or one drop) of developer A to each well, then add 50μl (or one drop) of developer B, and mix gently Evenly, develop color for 15 minutes in the dark at 37 ℃ 10. Termination: Add 50μl of stop solution (or one drop) to each well to stop the reaction (the blue color turns to yellow at this time). 11. Determination: Measure the absorbance (OD value) of each well in sequence with the blank air conditioner at zero and 450nm wavelength. Determination should be terminated Within 15 minutes after the solution. Test effectiveness: the average of positive control wells ≥1.00; the average of negative control wells ≤0.15 Calculation of critical value (CUT OFF): critical value = average value of negative control wells + 0.20 Negative judgment: the sample whose OD value is less than the critical value (CUT OFF) is negative for influenza A type IgM (FLU-A IgM) Positive judgment: the sample whose OD value ≥ critical value (CUT OFF) is positive for influenza A type IgM (FLU-A IgM) Precautions 1. The operation is strictly in accordance with the instructions. The components of different batches of this reagent must not be mixed. 2. The kit should be equilibrated at room temperature for 15-30 minutes before being taken out of the refrigerated environment. After use, the slats should be stored in sealed bags. 3. Crystals may be precipitated in the concentrated washing liquid, which can be heated and dissolved in a water bath during dilution, and the results will not be affected during washing. 4. The sealing film is limited to one-time use to avoid cross-contamination. 5. Please keep the substrate away from light. 6. The test results must be determined based on the reading of the microplate reader. When using dual wavelength detection, the reference wavelength is 630nm 7. All samples, washing liquids and various wastes should be treated as infectious agents. The stop solution is 2M sulfuric acid, which must be used be careful. 8. If there is any difference with the English manual, the English manual shall prevail .. specification: 96 servings / box Storage conditions and validity period 1. Kit storage :; 2-8 ℃. 2. Validity: 6 months J-Caine Lidocaine,J-Cain Numbing Cream Lidocaine,Glutathione Whitening Injection,J-Cain 15.6% Lidocaine Cream Shijiazhuang Asa Technology Co., Ltd. , https://www.hskinlift.com
This kit is for research use only.
Drug Name:
1 20 times concentrated washing solution 30ml × 1 bottle 7 stop solution 6ml × 1 bottle
2 Enzyme label reagent 6ml × 1 / bottle 8 Positive control 0.5ml × 1 vial
3 Enzyme label coated plate 12 well × 8 strips 9 Negative control 0.5ml × 1 bottle
4 Sample diluent 6ml × 1 bottle 10 Instructions 1 copy
5 Developer A solution 6ml × 1 bottle 11 sealing film 2 sheets
6 Developer B solution 6ml × 1 bottle 12 sealed bag 1
Specimen processing and requirements:
7. Incubation: The operation is the same as 3.
Result judgment: