Human anti-internal factor antibody (IFA) ELISA kit analysis and detection instructions This kit is for research use only. Detection range: 48T25ng/L-800ng/L purpose of usage: This kit is used to determine the anti-inner factor antibody (IFA) content of human serum, plasma and related liquid samples. Experimental principle The kit uses a double antibody sandwich assay to determine the level of human anti-in vivo factor antibody (IFA) in the specimen. The microplate was coated with purified human anti-in vivo antibody (IFA) antibody to prepare a solid phase antibody, and anti-internal factor antibody (IFA) was sequentially added to the microcapsule of the coated monoclonal antibody, followed by HRP-labeled anti-inner factor. The antibody (IFA) antibody binds to form an antibody-antigen-enzyme-labeled antibody complex, which is thoroughly washed and then added to the substrate TMB for color development. TMB is converted to blue under the catalysis of HRP enzyme and converted to the final yellow color by the action of an acid. The color depth is positively correlated with the anti-inner factor antibody (IFA) in the sample. The absorbance (OD value) was measured with a microplate reader at a wavelength of 450 nm, and the concentration of human anti-internal factor antibody (IFA) in the sample was calculated from a standard curve. Kit composition 1.30 times concentrated washing solution 20ml × 1 bottle; 2. enzyme standard reagent 6ml × 1 bottle 3. Enzyme label coating plate 12 holes × 8; 4. Sample dilution 6ml × 1 bottle 5. Color developer A liquid 6ml × 1 bottle; 6. Color developer B liquid 6ml × 1 bottle 7. Stop solution 6ml × 1 bottle; 8. Standard (48ng / ml) 0.5ml × 1 bottle 9. Standard dilution 1.5ml × 1 bottle; 10. Instructions 1 11. 2 sheets of sealing film; 12. 1 sealed bag 150 μl of Standard 2 is added to 150 μl of standard dilution 2. Loading: Set blank holes separately (the blank control wells are not added with the sample and the enzyme standard reagent, the other steps are the same), the standard holes, and the sample holes to be tested. Accurately load 50 μl of the standard on the enzyme-labeled plate, add 40 μl of the sample dilution to the well to be tested, and then add 10 μl of the sample to be tested (the final dilution of the sample is 5 times). Add the sample to the bottom of the well of the microplate, try not to touch the wall of the well, and shake gently to mix. 3. Incubation: The plate was sealed with a sealing film and incubated at 37 ° C for 30 minutes. 4. Dosing: 20 times concentrated washing solution diluted with distilled water 20 times and used 5. Washing: Carefully remove the sealing film, discard the liquid, dry it, fill each well with the washing liquid, let stand for 30 seconds, then discard it, repeat 5 times, and pat dry. 6. Add enzyme: 50 μl of enzyme labeling reagent was added to each well, except for blank wells. 7. Incubation: The operation is the same as 3. 8. Washing: The operation is the same as 5. 9. Color development: Add 50 μl of the developer to each well, then add 50 μl of the developer, gently shake and mix, and color at 37 ° C for 15 minutes. 10. Termination: 50 μl of stop solution was added to each well to terminate the reaction (when the blue color turned yellow). 11. Measurement: The absorbance (OD value) of each well was measured sequentially with a blank air conditioner of zero and a wavelength of 450 nm. The measurement should be carried out within 15 minutes after the addition of the stop solution. Advantages of ELISA kits: Comprehensive - a comprehensive screening of autoimmune diseases by mixing 8 different common antigens. Fast - short detection time (~2 hours), early diagnosis time. Accurate - ELISA detection method, high sensitivity, specificity, and good applicability. High quality - American technology development, high quality and high reliability analytical performance. Low price - domestic production, in line with domestic conditions, reducing production costs. Elisa kit organization structure: 1. Serum: Avoid any cell irritation during the procedure. Use tubes without pyrogens and endotoxins. After collecting the blood, the red blood cells were quickly and carefully separated by centrifugation at 1000 x g for 10 minutes. 2. Plasma: EDTA, citrate, heparin plasma can be used for detection. The pellet was removed by centrifugation at 1000 x g for 30 minutes. 3. Cell supernatant: Centrifuge at 1000 xg for 10 minutes to remove particles and polymer. 4, tissue homogenate: the tissue is added to the appropriate amount of physiological saline chopped. Centrifuge at 1000 xg for 10 minutes and take the supernatant. 5. Storage: If the sample is not used immediately, it should be divided into small parts - 70 ° C to avoid repeated freezing. Do not use hemolysis or hyperlipemia as much as possible. If there are large amounts of particles in the serum, centrifuge or filter before testing. Do not heat thaw at 37 ° C or higher. It should be thawed at room temperature and ensure that the sample is fully thawed evenly. Precautions: 1. The kit should be taken out from the refrigerated environment and should be used at room temperature for 15-30 minutes before use. If the enzyme label is unsealed after opening, the slats should be stored in a sealed bag. 2. Concentrated washing liquid may crystallize out. When diluted, it can be heated and dissolved in a water bath. The washing will not affect the result. 3. The sampler should be used for each step, and the accuracy should be corrected frequently to avoid test errors. The loading time is controlled within 5 minutes. If the number of specimens is large, it is recommended to use a gun. 4. Please make a standard curve at the same time for each measurement and make a double hole. If the content of the substance to be tested in the specimen is too high (the OD value of the sample is larger than the OD value of the standard pore), please first dilute the sample dilution with a certain multiple (n times) and then measure it. When calculating, multiply the total dilution factor ( ×n×5). 5. The sealing film is intended for single use only to avoid cross-contamination. 6. Please keep the substrate away from light. 7. Strictly follow the instructions of the manual, the test results must be based on the microplate reader reading. 8. All samples, washings and various wastes should be treated as infectious materials. 9. The different batch components of this reagent must not be mixed. 10. If it is different from the English manual, the English manual shall prevail. Storage conditions and expiration date 1. The kit is stored at: 2-8 °C. 2. Validity: 6 months 7 Inch Face Access Control,Face Recognition Access Control With Fingerprint,7 Inch Ultra Thin Face Recognition Terminal,Dual Camera Recognition Access Control System Shenzhen BIO Technology Co.,Ltd. , https://www.huifantech.com