Manual of Human Monoamine Oxidase (MAO) ELISA Kit

Human Monoamine Oxidase (MAO) Enzyme Linked Immunoassay (ELISA) Kit User Manual Xiamen Huijia Biotechnology Co., Ltd. This kit is for research use only. Drug name: Generic name: Human monoamine oxidase (MAO) enzyme-linked immunoassay kit Purpose of use: This kit is used to determine the content of monoamine oxidase A (MAO) in human serum, plasma and related liquid samples. Experimental principle The kit uses an indirect method to determine the level of human monoamine oxidase (MAO) in the specimen. A microplate was coated with purified human monoamine oxidase (MAO) antibody to make a solid-phase antibody. Monoamine oxidase A (MAO) standards of known concentration and monoamine oxidase A (MAO) of unknown concentration were added to the microwells coated with mAb in sequence. ) To be tested, after incubation, add biotin-labeled anti-IgG antibody, and then combine with streptavidin-HRP to form an immune complex. After thorough washing, 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. The color depth is positively correlated with the monoamine oxidase A (MAO) in the sample. The absorbance (OD value) was measured with a microplate reader at a wavelength of 450 nm, and the concentration of human monoamine oxidase (MAO) in the sample was calculated by a standard curve. Kit composition 1 20 times concentrated washing solution 50ml × 1 bottle 9 standard S1 (180 U / L) 0.5ml × 1 bottle 2 streptavidin-HRP 6ml × 1 bottle standard S2 (120 U / L) 0.5 ml × 1 bottle 3 enzyme-coated plates 12 wells × 8 standard S3 (60 U / L) 0.5ml × 1 bottle 4 biotinylated anti-IgG antibody 6ml × 1 standard S4 (30 U / L) ) 0.5ml × 1 bottle of 5 developer A solution 6ml × 1 bottle of standard product S5 (15U / L) 0.5ml × 1 bottle of 6 developer B solution 6ml × 1 / bottle 10 sealed bag 1 7 stop solution 6ml × 1 bottle of 11 sealing film 3 sheets 8 sample diluent 6ml × 1 bottle 12 instruction manual 1 specimen requirement 1. The samples containing NaN3 cannot be detected because NaN3 inhibits horseradish peroxidase (HRP) activity. 2. Specimens are extracted as soon as possible after collection, and extraction is performed according to relevant literature, and experiments should be conducted as soon as possible after extraction. If the test cannot be performed immediately, the specimen can be stored at -20 ° C, but repeated freeze-thaw operation steps should be avoided to determine the required number of slats based on the number of samples to be tested plus the number of standards. It is recommended to make multiple holes for each standard and blank hole. Each sample is determined according to its own quantity, and those that can use multiple holes can be used as much as possible. Sample addition: set up blank wells (the blank control wells do not add samples, the rest of the operation is the same), standard wells, sample wells to be tested. Then add 50μl of the standard to the standard well, add 10μl of the sample to be tested to the sample reaction well and then add 40μl of the sample diluent (the final dilution of the sample is 5 times), cover with the sealing film, shake gently and mix well, 37 ℃ Incubate for 45 minutes. Mixing solution: dilute 20 times concentrated washing solution with distilled water 20 times and then wash for standby: carefully remove the sealing film, discard the liquid, spin dry, fill each well with the washing solution, let it stand for 30 seconds and then discard, repeat 4 Time, pat dry. Add biotin-labeled anti-IgG antibody: Add 50 μl of biotin-labeled anti-IgG antibody to each well. Incubate at 37 ° C for 30 minutes to wash: operate the same as 4. Adding streptavidin-HRP: add 50 μl streptavidin-HRP to each well, gently shake and mix, and incubate at 37 ° C for 30 minutes. Washing: The operation is the same as 4. Color development: add 50μl of developer A to each well, then add 50μl of developer B, mix gently, and develop at 37 ° C in the dark for 15 minutes. Termination: add 50μl of stopper solution to each well to stop the reaction (at this time Blue to yellow). Measurement: The absorbance (OD value) of each well was measured in sequence with the blank air conditioner at zero and 450 nm wavelength. The measurement should be carried out within 15 minutes after adding the stop solution. Calculate the standard concentration as the abscissa and the OD value as the ordinate. Draw a standard curve on the coordinate paper, and find the corresponding concentration from the standard curve according to the OD value of the sample; multiply by the dilution factor; or use the standard The linear regression equation of the standard curve is calculated from the concentration and the OD value. The OD value of the sample is entered into the equation to calculate the sample concentration and then multiplied by the dilution factor, which is the actual concentration of the sample. Matters needing attention 1. The kit should be equilibrated at room temperature for 15-30 minutes before being taken out of the refrigerated environment. If the enzyme label coated plate is unopened, the strip should be stored in a sealed bag. 2. 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. 3. The sampler should be used at each step of sample addition, and the accuracy should be regularly checked to avoid test errors. It is best to control the sampling time within 5 minutes. If there are many specimens, it is recommended to use a volley gun to add samples. If the content of the substance to be tested in the specimen is too high (the OD value of the sample is greater than the OD value of the first well of the standard well), please dilute the sample by a certain multiple (n times) and then determine it. When calculating, please multiply the dilution factor (× 5 × n). The sealing film is limited to one-time use to avoid cross-contamination. 6. The components of different batches of this reagent shall not be mixed. Store developer B in dark place. 7. Strictly follow the instructions, and the test results must be determined based on the reading of the microplate reader. 8. All samples, washing liquids and various wastes should be treated as infectious agents. 9. If there is any difference with the English manual, the English manual shall prevail. Detection range: 0.9 IU / ml-60 IU / ml Sensitivity: 0.225 IU / ml Specification: 96 servings / box Storage conditions and expiration date Kit storage :; 2-8 ℃. 2. Validity: 6 months

Storage Rack

Storage Rack,Living Room Snack Racks,3 Floors Plastic Shelving,Multilayer Storage Rack

Taizhou Baiying Commdity Co., Ltd , https://www.baiyingplastics.com