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Human 12-hydroxyeicosatetraenoic acid (12-HETE) elisa kit Instructions for use 1. Dilution and loading of standards: Set standard wells on the enzyme label coated plate, 10 holes, in the first, the first Add 100 μl of the standard to the two wells, then add 50 μl of the standard dilution to the first and second wells and mix; then add 100 μl from each of the first and second wells to the third and fourth. Holes, then add 50 μl of the standard dilution solution to the third and fourth wells, and mix; then discard 50 μl in each of the third and fourth wells, and then add 50 μl to each of the fifth and sixth wells. Then, add 50 ul of the standard dilution solution in the fifth and sixth holes, and mix; after mixing, take 50 μl from each of the fifth and sixth holes and add them to the seventh and eighth holes, respectively. Add 50 μl of the standard dilution solution to the eighth well, mix and mix 50 μl from the seventh and eighth wells into the ninth and tenth holes, and add the standard dilution in the ninth and tenth holes respectively. 50 μl, after mixing, take 50 μl from each of the ninth and tenth holes and discard. (The amount of each well was 50 μl after dilution, and the concentrations were 1800 ng/L, 1200 ng/L, 600 ng/L, 300 ng/L, 150 ng/L, respectively). 2. Loading: Set blank holes separately (the blank control wells are not added with the sample and the enzyme standard reagent, the other steps are the same), and the sample holes to be tested. Add 40 μl of the sample dilution to the sample well to be tested on the enzyme-labeled plate, and then add 10 μl of the sample to be tested (the final dilution of the sample is 5 times). Add the sample to the bottom of the well of the microplate, try not to touch the wall of the well, and shake gently to mix. 3. Incubation: The plate was sealed with a sealing film and incubated at 37 ° C for 30 minutes. 4. Solution: 30 (48 times of 20T) concentrated washing solution was diluted with distilled water 30 (20 times of 48T) and used. 5. Washing: Carefully remove the sealing film, discard the liquid, dry it, fill each well with the washing solution, let stand for 30 seconds, then discard it, repeat 5 times, and pat dry. 6. Add enzyme: 50 μl of enzyme labeling reagent was added to each well, except for blank wells. 7. Incubation: operation is the same as 3. 8. Washing: operation is the same as 5. 9. Color: first add coloring agent A50μl to each well, then add coloring agent B50μl, gently shake and mix, avoid light at 37 °C 15 minutes. 10. Termination: 50 μl of stop solution was added to each well to terminate the reaction (when the blue color turned yellow). 11. Measurement: The absorbance (OD value) of each well was measured sequentially with a blank air conditioner of zero and a wavelength of 450 nm. The measurement should be carried out within 15 minutes after the addition of the stop solution. [Human 12-hydroxyeicosatetraenoic acid (12-HETE) elisa kit] Note: 1. The kit should be taken out from the refrigerated environment and allowed to equilibrate for 15-30 minutes at room temperature. If the enzyme label is unsealed after unsealing, the strip should be stored in a sealed bag. 2. Concentrated washing liquid may crystallize out. When diluted, it can be heated and dissolved in a water bath. The washing does not affect the result. 3. The sampler should be used for each step and the accuracy should be corrected frequently to avoid test errors. It is best to control the loading time within 5 minutes. If the number of specimens is large, it is recommended to use a gun. 4. Please make a standard curve at the same time of each measurement, it is best to make a double hole. If the content of the substance to be tested in the specimen is too high (the OD value of the sample is larger than the OD value of the first hole of the standard well), please first dilute the sample dilution with a certain multiple (n times) and then measure it. When calculating, multiply the total dilution by the total dilution. Multiple (×n×5). 5. The sealing film is intended for single use only to avoid cross-contamination. 6. Keep the substrate away from light. 7. Strictly follow the instructions of the manual, the test results must be based on the microplate reader reading. 8. All samples, washings and various wastes should be treated as infectious materials. 9. The different batch components of this reagent must not be mixed. 10. In the case of an English manual, the English manual shall prevail. Kit performance: 1. Sample linear regression and expected concentration correlation coefficient R value of 0.95 or more. 2. Within and within the batch should be less than 9% and 11% detection range: 0.2IU / L - 6IU / L of the company's product quality assurance, for the domestic excellent ELISA kit suppliers, the price is fair, and provide you with integrity Reliable service, welcome to inquire product description and other details, 60484409
April 08, 2021