Instructions for human immunoglobulin G detection ELISA kit Shanghai Qiao Yu ELISA kit supplier! This reagent is for research use only. Specimen: serum or plasma Note: This product is for scientific research only! Human immunoglobulin G detection ELISA kit test principle: The IGG kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA). Standards of known IGG concentration and samples of unknown concentration are added to the microplates for detection. IGG and biotinylated antibodies are first incubated simultaneously. After washing, avidin-labeled HRP was added. After incubation and washing, the unbound enzyme conjugate is removed, then the substrates A, B, and the enzyme conjugate are added simultaneously. Produce color. The depth of the color is proportional to the concentration of IGG in the sample. Sample collection: 1, treatment and preservation methods: serum ... avoid any cell stimulation during the operation. 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 testing. 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 homogenization... Add the appropriate amount of normal saline to the tissue. Centrifuge at 1000 xg for 10 minutes and take the supernatant. 5. Save... 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. Human immunoglobulin G detection ELISA kit operation notes: ◠Reagents should be stored according to the label instructions and returned to room temperature before use. Standards after dilution should be discarded and cannot be stored. ◠The slats not used in the experiment should be immediately put back into the bag and sealed to prevent deterioration. ◠Other reagents that are not used should be packaged or covered. Do not mix reagents of different batches. Use before warranty. ◠Use disposable tips to avoid cross-contamination. Avoid pipettes with metal parts when drawing stop solution and substrate A and B. ◠Use a clean plastic container to configure the wash solution. Mix all the ingredients and samples in the kit thoroughly before use. ◠Substrate A should be volatilized to avoid opening the lid for a long time. Substrate B is sensitive to light and avoids prolonged exposure to light. Avoid contact with hands and be toxic. The OD value should be read immediately after the experiment is completed. ◠The order of adding reagents should be the same to ensure that all wells are incubated for the same time. ◠Carry out the incubation according to the time indicated in the instruction manual, the amount of liquid addition and the order. Kit performance: Sensitivity: The minimum detection concentration is less than the No. 1 standard. The linearity of the dilution. The linear regression coefficient of the sample and the expected concentration correlation coefficient R value was 0.990. Specificity: Does not react with other cytokines. Repeatability: The coefficient of variation between the plates and the plates is less than 10%. Human immunoglobulin G detection ELISA kit steps: Mix all reagents thoroughly before use. Do not allow the liquid to generate a large amount of foam, so as to avoid adding a large amount of air bubbles during the loading, resulting in errors in the loading. The number of slats required is determined by the number of samples to be tested plus the number of standards. Multiple holes are recommended for each standard and blank hole. Each sample is determined according to its own quantity, and it is possible to use the double hole as much as possible to make a double hole. 50 ul of the diluted standard was added to the reaction well, and 50 ul of the sample to be tested was added to the reaction well. Immediately add 50 ul of biotinylated antibody. Cover the membrane, mix gently by shaking, and incubate for 45 minutes at 37 °C. Remove the liquid from the well, fill each well with the washing solution, shake for 30 seconds, remove the washing solution, and pat dry with absorbent paper. Repeat this operation 4 times. If washing with a washer, the number of washings is increased once. 100 ul of affinity streptavidin-HRP was added to each well, and the mixture was gently shaken and incubated at 37 ° C for 30 minutes. Remove the liquid from the well, fill each well with the washing solution, shake for 30 seconds, remove the washing solution, and pat dry with absorbent paper. Repeat this operation 4 times. If washing with a washer, the number of washings is increased once. 50 μl of each of the substrates A and B was added to each well, and the mixture was gently shaken and incubated at 37 ° C for 5 minutes. Avoid lighting. Remove the microplate and quickly add 50 ul of stop solution. Immediately after adding the stop solution, the results should be determined. Limit The results above the No. 6 standard are non-linear and no accurate results can be obtained from this standard curve. Kit performance 1. Sensitivity: The minimum detection concentration is less than the No. 1 standard. The linearity of the dilution. The linear regression coefficient of the sample and the expected concentration correlation coefficient R value was 0.990. 2. Specificity: Does not react with other cytokines. 3. Repeatability: The coefficient of variation between the plate and the plate is less than 10%. Result judgment and analysis 1. Instrument value: read the OD value of each hole on a microplate reader with a wavelength of 450 nm. 2. The absorbance OD value is the ordinate (Y), and the corresponding Amylin standard concentration is the abscissa (X), and the corresponding curve is obtained. The Amylin content of the sample can be converted into the corresponding concentration according to the OD value from the standard curve. 3. Detection range: 0-80ng/ml 4. Sensitivity: 0.1 ng/ml Nutrition Biscuit,Black Tartary Buckwheat Sachima,Health Nutritious Tartary Buckwheat Twist,Xylitol Sesame Buckwheat Crisp Huantai Biotechnology Co., Ltd. , https://www.huantaifds.com