Recently, technical experts from Shanghai Hengyuan Experimental Department contacted Manager He to communicate the latest Elisa technology. According to Mr. He, the major schools have started the summer vacation so far, and Shanghai Hengyuan biotech technicians have compiled a summary of the operational problems common to customers since this semester. The following is a technical expert analysis of the Elisa kit test report:
1. The so-called single-wavelength colorimetric is equal to the usual colorimetric determination at a wavelength having a maximum absorption for color development such as 450 nm or 492 nm;
2. For dual-wavelength two-color, the microplate reader is measured at sensitive wavelengths such as 450 nm and non-sensitive wavelengths of 630 nm. The absorbance of the sensitive wave is measured by the absorbance of the specific color of the enzyme reaction and the fingerprint and scratch on the plate. The sum of absorbance caused by dirt such as dust;
3. The measurement at the non-sensitive wavelength changes the wavelength to a certain value. The Elisa kit makes the absorbance value of the specific color development of the sample reaction to be zero, and the absorbance measured at this time is the absorbance value of the dirt.
TMB is used as a substrate and an OPD-based kit is used. The former has a colorimetric wavelength of 450 nm and the latter has a wavelength of 492 nm. The filter needs to be replaced at any time according to requirements. Finally, the value given by the microplate reader is the difference between the absorbance value at the sensitive wavelength and the absorbance value at the extreme sense wavelength. For the test with soft board as the carrier, the board needs to be placed in a 96-hole frame for colorimetry. When the Elisa kit is colorimetric, the substrate should be calibrated with distilled water to read the substrate well.
Because there is a certain degree of uncertainty in the non-specific absorption of individual vacant wells in the Elisa kit assay, that is, each absorbance or the same position of the vacant wells may result in different absorbance measurements, so it is determined by Elisa. When colorimetric, it is best to use dual wavelength colorimetry. Therefore, the dual-wavelength colorimetric measurement has the advantage of eliminating the influence of non-specific absorption of the microtiter plate itself, the non-specific absorption of the specimen in the plate hole, fingerprints, scratches, dust, and the like on the specific colorimetric measurement of the absorbance.
In addition, Shanghai Hengyuan Bio Company reminds everyone that during the operation,
In the Elisa experimental measurement, it should also be noted that the samples before and after dilution must be thoroughly mixed, and all reagents should be shaken before loading to ensure the uniformity of the test. To learn more about Elisa technology, you are welcome to visit Shanghai Hengyuan website for the latest product catalogs and operating technologies.
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