Spot human glycated albumin ELISA detection kit
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Human glycated albumin ELISA detection kit This reagent is for research use only Specimen: Serum or plasma Test principle: The GA kit is a solid-phase sandwich method enzyme-linked immunosorbent assay (ELISA). Standards with known GA concentrations, unknown concentrations The sample is added to the microplate for detection. First, GA and biotin-labeled antibodies are incubated simultaneously. 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 GA in the sample. Bring your own material distilled water. Sampler: 5ul, 10ul, 50ul, 100ul, 200ul, 500ul, 1000ul. Oscillator and magnetic stirrer etc. 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. Handling precautions 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. When washing the enzyme-labeled plate, it should be fully patted dry. 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. Human glycated albumin ELISA detection kit 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 cell supernatant 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. Reagent preparation standards: Serial dilutions of standards should be prepared during the experiment and cannot be stored. Before dilution, the standard was mixed by shaking. Dilution of washing buffer (50 ×): 50-fold dilution with distilled water. 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. The specimen was diluted 1: 1 with the specimen dilution solution and then added 50ul to the reaction well. 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, gently shake and mix, and incubate at 37 ° C for 1 hour. 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 3 times. If washing with a plate washer, the number of washes is increased once. Add 80 ul of streptavidin-HRP to each well, gently shake and 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 3 times. If washing with a plate washer, the number of washes is increased once. Add 50ul of substrate A and B to each well, mix gently by shaking, and incubate at 37 ° C for 10 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. The results of the human glycated albumin ELISA detection kit limited to No. 6 standard and above are non-linear, and accurate results cannot be obtained according to this standard curve. Kit performance 1. Sensitivity: The minimum 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%. Judgment and analysis of results 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), and the corresponding GA standard concentration as the abscissa (X) To make the corresponding curve, the GA content of the sample can be converted to the corresponding concentration from the standard curve according to its OD value. 3. Detection value range: 0-80mmol / L4, sensitivity: 0.1 mmol / L human glycated albumin ELISA detection kit DA2315-0.2ml LRIG3 (leucine rich repeats and immunoglobulin-like domains 3) anti-LRIG3 antibody 0.2ml DA002-2ml human CD4 Monoclonal antibody / beads anti-human CD4 monoclonal antibody immunomagnetic beads 2mlDA003-2ml human CD8 Monoclonal antibody / beads anti-human CD8 monoclonal antibody immunomagnetic beads 2mlDA2316-0.1ml LRP / MVP (Lung resistance related protein) 0.1mlDA2316-0.2ml LRP / MVP (Lung resistance related protein) Lung resistance related protein antibody 0.2mlDA2317-0.2ml LRRC4 (Leucine-rich repeat-containing protein 4) Brain tumor related gene antibody 0.2mlDA2318-0.1ml LRRK2 (Leucine- rich repeat kinase 2) Leucine-rich repeat kinase 2 antibody 0.1mlDA2318-0.2ml LRRK2 (Leucine-rich repeat kinase 2) Leucine-rich repeat kinase 2 antibody 0.2mlDA2319-0.1ml Iumbrokinase earthworm plasmin antibody / anti-worm kinase Antibody 0.1mlDA2319-0.2ml Iumbrokinase Earthworm Fibrinolytic Antibody / Anti-Lumkinase Antibody 0.2ml DA2372-0.2ml Megsin / SER—PINB7 Serine (or Cysteine) Protein Inhibitor B7 antibody 0.2ml DA2376-0.2ml M-enk anti-methionine enkephalin antibody 0.2ml DAR1143-0.1ml Rabbit anti-MK IgM / APC? Fluorescein APC labeled rabbit anti-monkey IgM 0.1ml DAR1144-0.1ml Rabbit anti- pig IgG / APC fluorescein APC labeled rabbit anti-pig IgG 0.1ml DA2320-0.2ml Lxn (Latexin) carboxypeptidase inhibitor antibody 0.2ml DA2321-0.2ml Ly-6G (Lymphocyte antigen 6G) lymphocyte antigen 6G antibody 0.2ml