Shanghai Fusheng Industrial Co., Ltd. specializes in supplying ELISA kits, kits, detection kits, human ELISA kits, rat ELISA kits, mouse ELISA kits, ELISA kits, ELISA KIT, rabbit ELISA kits, guinea pig ELISA Kit, cattle ELISA kit, sheep ELISA kit, dog ELISA kit, pig ELISA kit, hamster ELISA kit, kit, detection reagents, antibodies, cell lines, culture medium, tumor necrosis factor, interleukin 2, interference Factors, standards, controls, biological reagents, markers, inhibitors, antigens, agar, cell growth factors, etc. For details, please call the contact person: Manager Zhang telephone and fax. Human secreted immunoglobulin AELISA detection kit test principle: sIgA kit is a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA). Standards with known sIgA concentration and samples with unknown concentration are added to microwell enzyme plates Detection. First, sIgA and biotin-labeled antibody were incubated at the same time. After washing, HRP labeled with avidin was added. After incubation and washing, the unbound enzyme conjugate is removed, and then substrates A and B are added to act simultaneously with the enzyme conjugate. Produce colors. The color depth is proportional to the concentration of sIgA in the sample. Bring your own material: distilled water. Sampler: 5ul, 10ul, 50ul, 100ul, 200, 500ul, 1000ul. Oscillator and magnetic stirrer etc. Sample collection, processing and storage methods: serum-avoid any cell stimulation during the operation. Use test tubes free of pyrogens and endotoxins. After collecting blood, centrifuge at 1000 × g for 10 minutes to separate the serum and red blood cells quickly and carefully. Plasma ----- EDTA, citrate, heparin plasma can be used for detection. Centrifuge at 1000 × g for 30 minutes to remove particles. The supernatant of the cells was centrifuged at 1000 × g for 10 minutes to remove particles and polymers. Tissue homogenate ----- Mash the tissue with appropriate amount of saline. Centrifuge at 1000 × g for 10 minutes, and take the supernatant for storage. If the sample is not used immediately, it should be divided into small parts and stored at -70 ℃ to avoid repeated freezing. If possible, do not use hemolysis or hyperlipidemia. If there are a large number of particles in the serum, centrifuge or filter before testing. Do not thaw at 37 ° C or higher. Thaw at room temperature and ensure that the sample is thawed evenly and adequately. Precautions for operation of human secreted immunoglobulin AELISA detection kit: reagents should be stored according to label instructions, and returned to room temperature before use. The sparse standards should be discarded and cannot be stored. The slats not used in the experiment should be immediately returned to the packaging bag, sealed and stored to avoid deterioration. Unused other reagents should be packed or covered. Do not mix reagents of different batches. Use before warranty. Use a disposable pipette tip to avoid cross-contamination, and avoid using a sampler with a metal part when drawing the stop solution and substrates A and B. Use a clean plastic container to prepare the washing solution. Mix all components and samples in the kit thoroughly before use. Substrate A should evaporate to avoid opening the lid for a long time. Substrate B is sensitive to light and avoid prolonged exposure to light. Avoid contact with hands, it is 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. Perform the incubation operation according to the time, the amount and order of the liquid indicated in the instructions. Safety: Avoid direct contact with the stop solution and substrates A and B. Once exposed to these liquids, rinse with water as soon as possible. Do not eat, drink, smoke or use cosmetics during the experiment. Do not use your mouth to absorb any ingredients in the kit. Kit performance: 1. Sensitivity: The smallest detection concentration is less than No. 1 standard. Linearity of dilution. The correlation coefficient between the linear regression of the sample and the expected concentration is 0.990. 2. Specificity: Does not react with other cytokines. 3. Repeatability: The coefficients of variation within and between plates are less than 10%. Human secreted immunoglobulin AELISA detection kit operation steps: before use, mix all reagents thoroughly. Don't make the liquid generate a lot of foam, so as to avoid adding a large number of bubbles during sample addition, which will cause errors in sample addition. The number of slats required is determined by the number of samples to be tested plus the number of standard products. 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. Add 50ul of the diluted standard to the reaction well and 50ul of the sample to be tested in the reaction well. Immediately add 50ul of biotin-labeled antibody. Cover the membrane plate, mix gently by shaking, and incubate at 37 ° C for 45 minutes. Shake off the liquid in the wells, fill each well with washing liquid, shake for 30 seconds, shake off the washing liquid, pat dry with absorbent paper. Repeat this operation 4 times. If washing with a plate washer, the number of washes is increased once. Add 100ul of streptavidin-HRP to each well, gently shake to mix, and incubate at 37 ° C for 30 minutes. Shake off the liquid in the wells, fill each well with washing liquid, shake for 30 seconds, shake off the washing liquid, pat dry with absorbent paper. Repeat this operation 4 times. If washing with a plate washer, the number of washes is increased once. Add 50ul each of substrate A and B to each well, gently shake to mix, and incubate at 37 ° C for 5 minutes. Avoid light. Remove the enzyme labeling plate and quickly add 50ul of stop solution. After adding the stop solution, the results should be measured immediately. The OD value of each well was measured at a wavelength of 450 nm. Judgment and analysis of the results of human secretory immunoglobulin AELISA detection kit: 1. Instrument value: read the OD value of each well on a microplate reader with a wavelength of 450 nm. 2. Take the absorbance OD value as the ordinate (Y), corresponding The concentration of the sIgA standard product is the abscissa (X), and the corresponding curve is made. The sIgA content of the sample can be converted from the standard curve to the corresponding concentration according to its OD value, and then multiplied by the dilution factor; or the concentration and OD value of the standard Calculate the regression equation of the standard curve, substitute the OD value of the sample into the equation, calculate the sample concentration, and multiply it by the dilution factor to get the actual concentration of the sample. 3. Detection value range: 0-40umol / L4, sensitivity: 0.1 umol / LPhospho-FADD (Ser191)? Phospho-MAPKAPK2 (Thr222)? Phospho-MAPKAPK2 (Thr222)? Phospho-mitogen-activated protein kinase Activated protein kinase 2 antibody 0.1ml phospho-ASK1 (Ser967)? Phosphorylation apoptosis signal regulating kinase 1 antibody 0.1ml phosphoM-CSF Receptor (Tyr809)? 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