Focus on the sample processing and requirements of ELISA Kit

This paper mainly introduces the sample processing of ELISA kit and the requirements of ELISA kit products The advantages and disadvantages of ELISA kit products are mainly judged from its specificity, accuracy, degree and other aspects Due to the space relationship, the specificity of products is explained in detail: the specificity refers to the specific amplification of the target segment rather than the performance of other segments in the PCR amplification process When the PCR experiment itself has high sensitivity and the experimental conditions are not fully optimized, there are usually other heterobands, that is, non-specific amplification The specificity of PCR amplification can be affected by the control of template, primer quality and reaction conditions In recent years, the research results show that the quality of buffer (such as ion type and composition, reaction optimizer, etc.) plays an important role in ensuring PCR specific amplification In general buffer, the salt that regulates hydrogen bond is only KCl, while the sensitivity and specificity of the buffer system of Dongsheng HSTMTaqMix through reaction optimization agent and KCl/ (NH4) S PCR is the characteristic of this one This also determines that our experimental system regulation is actually the balance of sensitivity and specificity suitable for experiments Because of many components in PCR system, there are many combinations and complicated screening work Dongsheng has designed pcrmix and hstaqmix respectively based on sensitivity and specificity to help you determine a large balance point If you need a more detailed balance point, please select the corresponding MixKit series for fine tuning The sample processing and requirements in the operation experiment of ELISA: 1 Serum: natural coagulation of blood at room temperature for 10-20 minutes, centrifugation for about 20 minutes (2000-3000 RPM) The supernatant should be collected carefully, and centrifugation should be carried out again in case of precipitation during storage 2 Plasma: EDTA or sodium citrate should be selected as anticoagulant according to the requirements of the sample After mixing for 10-20 minutes, centrifuge for about 20 minutes (2000-3000 RPM) The supernatant should be collected carefully, and centrifugation should be performed again in case of precipitation formation during preservation 3 Urine: collected with sterile tube and centrifuged for about 20 minutes (2000-3000 RPM) The supernatant should be collected carefully, and centrifugation should be performed again in case of precipitation formation during preservation Hydrothorax, ascites and cerebrospinal fluid were carried out by reference 4 Cell culture supernatant: collect the secretory components with sterile tube Centrifugation for about 20 minutes (2000-3000 RPM) Collect the supernatant carefully When detecting the components in cells, PBS (ph7.2-7.4) was used to dilute the cell suspension, and the cell concentration was about 1 million / ml Through repeated freezing and thawing, the cells can be destroyed and the intracellular components can be released Centrifugation for about 20 minutes (2000-3000 RPM) Collect the supernatant carefully In case of precipitation formation during storage, centrifugation shall be carried out again 5 Tissue specimen: after cutting the specimen, weigh it Add a certain amount of PBS, pH7.4 Refrigerate quickly with liquid nitrogen The temperature of the sample remained at 2-8 ℃ after melting Add a certain amount of PBS (pH7.4), and use hand or homogenizer to homogenize the sample fully Centrifugation for about 20 minutes (2000-3000 RPM) Collect the supernatant carefully One is to be tested after subpackage, and the rest is to be frozen for standby 6 The samples should be extracted as early as possible after collection, according to the relevant literature, and the experiment should be carried out as soon as possible after extraction If the test can not be carried out immediately, the samples can be stored at - 20 ℃, but repeated freezing and thawing should be avoided 7 The samples containing NaN3 can not be detected because of the inhibition of horseradish peroxidase (HRP) activity by NaN3.