The so-called single-wavelength colorimetric is equal to the usual colorimetric measurement at the wavelength with the largest absorption of color development, such as 450nm or 492nm; while the dual-wavelength dual-color is measured once at a sensitive wavelength such as 450nm and a non-sensitive wavelength 630nm The absorbance measured above and below the sensitive wave is the sum of the absorbance caused by the specific color development of the enzyme reaction of the sample and the absorbance caused by fingerprints, scratches, dust and other dirt on the plate well; measurement at non-sensitive wavelengths changes the wavelength to a certain value, EELISA The kit allows the sample to determine the absorbance value of the enzyme reaction specific color development to zero, and the measured absorbance at this time is the absorbance value of the dirt.
Both TMB as a substrate and OPD as a substrate are used. The colorimetric wavelength of the former is 450nm, and the latter is 492nm. 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 very sensitive wavelength. For the test with the soft board as the carrier, the board needs to be placed in a 96-well standard frame before color comparison. When the ELISA kit is used for color comparison, the zero point should be calibrated with distilled water and the substrate hole should be measured.
Because there is a certain degree of uncertainty in the non-specific absorption of a single vacancy hole in the ELISA kit measurement, that is to say, each measurement or the same measurement of the position of the vacancy hole may have different absorbance measurements, so in the ELISA measurement For colorimetry, it is best to use dual-wavelength colorimetry. ELISA kit Therefore, the dual-wavelength colorimetric measurement has the advantage that it can exclude the non-specific absorption, fingerprints, scratches, dust, etc. of the microtiter plate itself, the specimens in the wells of the plate, etc., on the specific color measurement absorbance. The expression of colorimetric results used to be the general optical density (OD), which is now defined as absorbance (A). Both have the same meaning.
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