ELISA standard curve

Generally speaking, for linear regression, the R2 of each test is best to reach 0.98, that is, R reaches 0.99, otherwise retest is required. If you need to do several inspections, the difference between slope and intercept should not exceed 20% each time, R is the correlation coefficient, and the correlation between x and y. It is generally represented by R2, which is more accurate. R2 is not said to be independent, the closer the value is to 1, the greater the correlation. However, the general R2 cannot reach this value. 0.95 does not necessarily mean that the correlation is low. It should also detect significant differences in correlation. If there is no significant difference, then R is credible. Of course, R2 is also credible. What is competitive ELISA? Competitive ELISA is based on the competitive binding technology, that is, the test molecule in the sample and a fixed amount of the same molecule (we generally use alkaline phosphatase as the label) and the same binding of the antibody immobilized on the well plate The sites compete, referring to the standard curve, and the test data values ​​are quantitative. What is sandwich ELISA? Sandwich ELISA uses an antibody specific for the analyte in the sample as the immobilized capture antibody, and then uses the second labeled antibody for detection. The detection antibody is also specific for the analyte. When referring to the standard curve, the test data values ​​are quantitative. How many samples can be made on a microplate is based on a 96-well microplate test. In addition to the positive control, negative control, background (blank) and detection antibody control, there are still 88 wells, which can be used for 44 samples (each sample is made in duplicate). Specimen collection • Serum: After the blood has naturally coagulated at room temperature for 10-20 minutes, centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant. If a precipitate forms, it should be centrifuged again. ◇ Plasma: EDTA, sodium citrate or heparin should be selected as anticoagulant according to the requirements of the kit, and 10% (v / v) anticoagulant should be added (0.1M sodium citrate or 1% heparin or 2.0% EDTA.Na2) After mixing for 10-20 minutes, centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If a precipitate forms, it should be centrifuged again. ◇ Urine, pleural and ascites fluid, cerebrospinal fluid: collected with sterile tubes. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If a precipitate forms, it should be centrifuged again. ◇ Cell culture supernatant: collect the sterile components when detecting secreted components. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. When detecting the components in the cells, dilute the cell suspension with PBS (PH7.2-7.4), and the cell concentration will reach about 1 million / ml. Through repeated freezing and thawing, the cells are destroyed and the intracellular components are released. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If a precipitate forms during storage, it should be centrifuged again. ◇ Organized specimen: weigh the weight after cutting the specimen. Add a certain amount of PBS, and 1 μg / L protease inhibitor or 50U / ml Aprotinin (aprotinin) can be added to the buffer. Use a hand or homogenizer to homogenize the specimen. Centrifuge for about 20 minutes (2000-3000 rpm). Carefully collect the supernatant and store at -20 degrees or -70 degrees. If necessary, the sample can be concentrated and dried. After aliquoting, a portion is to be tested, and the rest is frozen for future use.

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