plz anyone tel me ''In MTT Assay why do we measure OD at two differnt wave lengths ,,
i just knw 570 nm for test and 630 nm for reference.. anyone xplain me clearly plz
the two wavelengths measure the absorbance of the dye and of the background.
Which wave length is better for test, 490nm or 570nm.
In MTT assay we need to take o.D at two wave length because MTT colure will measure at 570 nm and turbidity of the cells we can measure at 630 nm. So for the clear dose response we need to take reading at both the wave length and 630 readings need to substract from 530 and than need to plote the reading. Which will give clear picture of the assay.
490 nm is used for MTS (red colour upon reduction) and 570 nm is used for MTT (blue colour in reduced form). so selection of wavelength depends upon the colour produced upon reduction of dye.