Spot human adrenal myelin ELISA test kit

 Human adrenal myelin ELISA test kit test principle: ADM kit is a solid-phase sandwich enzyme-linked immunosorption experiment (ELISA) The standard product of known ADM concentration, unknown concentration of samples added to the microporous enzyme plate for testing First, the antibodies of ADM and biotin markers are fed at the same time After washing, add the hrP labeled in affinity Then after temperature breeding and washing, remove the unbound enzyme bindings, and then add the substrate Sa, B, and the enzyme binding syllables simultaneously Produces color The depth of color is proportional to the concentration of ADM in the sample Distilled water from their own materials Samplers: 5ul, 10ul, 50ul, 100ul, 200ul, 500ul, 1000ul Oscillators and magnetic agitators, etc Safety avoids direct contact with terminating fluids 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 in the experiment Do not use your mouth to absorb any ingredients from the kit Human adrenal myelin ELISA test kit operating precautions reagents should be stored according to the label instructions, before use to restore to room temperature Standard items after dilute should be discarded and not kept The slats not used in the experiment should be immediately put back into the bag and sealed to avoid deterioration Other reagents that are not used should be packed or covered Reagents with different lot numbers should not be mixed Pre-use Use disposable tips to avoid cross contamination, and avoid using a sampler with metal parts when absorbing the termination fluid and the base A and B liquid Use a clean plastic container to configure the washing liquid Fullmix of ingredients and samples in the kit before use Wash the enzyme plate should be fully patted dry, do not put the absorbent paper directly into the enzyme label reaction hole to absorb water Base A should be volatile to avoid opening the lid for a long time Base B is sensitive to light and avoids prolonged exposure to light Avoid hand contact, toxic The OD value should be read immediately after the experiment is complete The order of the reagents added should be consistent to ensure that all reaction plates are fed at the same time In accordance with the specification indicated in the time, the amount of fluid and the order of the temperature breeding operation Sample collection, processing and preservation method serum ----- avoid any cell irritation during operation Use test tubes that do not contain hot and endotoxins After the blood is collected, 1000 x g centrifugated for 10 minutes the serum and red blood cells are quickly and carefully separated Plasma ----- EDTA, citrate, heparin plasma can be used for detection 1000 x g centrifugation for 30 minutes to remove the particles Cell on-the-top liquid --- 1000 x g centrifugation for 10 minutes to remove particles and polymers Save ------ If the sample is not used immediately, it should be stored in small portions - 70 degrees C to avoid repeated freezing Do not use hemolytic or hyperlipidemia whenever possible If there are a large number of particles in the serum, centrifugal or filtered before detection Do not heat the thaw at 37 degrees C or higher Thaw at room temperature and ensure that the sample is evenly and fully thawed The human adrenal myelin ELISA test kit is fully mixed with all reagents before the procedure is used Do not make the liquid produce a large amount of foam, so as not to add a large number of bubbles when adding, resulting in a sample addition error The number of slats required is determined based on the number of samples to be tested plus the number of standard strains Each standard and blank hole is recommended for compound holes Each sample according to its own quantity to determine, can use the compound hole as far as possible to do the compound hole Add diluted standard 50ul in the reaction hole, add the sample to be tested 50ul in the reaction hole Immediately add 50ul biotin-labeled antibodies Cover the membrane plate, gently oscillate and mix, 37 degrees C temperature for 1 hour Shake off the liquid in the hole, fill each hole with washing liquid, oscillate for 30 seconds, shake off the washing liquid, and beat dry with absorbent paper Repeat this 3 times If washing with a washing machine, the number of washing times increases once Add 80ul of affinity streptose-HRP to each hole, gently oscillate and mix, 37 degrees C temperature for 30 minutes Shake off the liquid in the hole, fill each hole with washing liquid, oscillate for 30 seconds, shake off the washing liquid, and beat dry with absorbent paper Repeat this 3 times If washing with a washing machine, the number of washing times increases once Each hole adds the substrate A, B 50ul each, gently oscillates mixed, 37 degrees C temperature for 10 minutes Avoid lighting Remove the enzyme plate, quickly add 50ul termination liquid, add the termination liquid should be immediately determined results The OD values of each hole are measured at a wavelength of 450nm The results above standard No 6 are non-linear, and the results cannot be obtained according to this standard curve Human adrenal myelin ELISA test kit performance 1 Sensitivity: Zui small detection concentration is less than Standard No 1 Linearity of dilution The sample linear regression coefficient r-factor associated with the expected concentration is 0.990 2 Specificity: Does not react with other cytokines 3 Repeatability: In- and inter-board variation coefficients are less than 10% Results And analysis 1, instrument value: on the wavelength 450nm enzyme labeling instrument to read the OD value of each hole 2, to absorbthe OD value of the ordinate (Y), the corresponding ADM standard concentration is horizontal coordinates (X), do the corresponding curve, the sample ADM content can be converted from the standard curve according to its OD value 3, detection value range: 0-400pg/ml4, sensitivity: 1.0 pg/mlNK-2R (Neurokinin A eit; SKR; Subtance-K receptor) Neurogenic A receptor antibody 0.2mlNKA (Neurokinin A) neurokinin A antibody 0.2mlStreptavidin/PE fluorofluorometer PE marker streptomyc incoc incogen 0.1mlAvidin/PE fluorofluorin PE labeled affinity 0.1mlrec Protein A/PE fluorolastatin PE labelre purified protein A 0.1ml human CD16 monoclonal anti-human CD16 monoantigen 100Thuman CD25 Monoclonal anti-body/FITC fluorolastin FITC labeled anti-human CD25 monoantigen 100T