I use Elisa micro plate reader which supports 96-well plate. problem is my absorbance values are almost same for standards. I use Methanol as solvent with Paclitaxel at 230nm even i change wavelength but got almost same absorbance values can anyone guide me ?
The plate is made up of polystyrene which is not suitable to measure absorbance at 230nm i.e. in UV range hence all measured absorbance values are same. Better take absorbance by using a quartz cuvette at 230nm.
Even glass and plastic cuvettes are be never used for absorbance measurements in UV range i.e. 200 to 380 nm.
You can try to mesure using Corning® 96-well UV-Transparent Microplates (the mean background O.D. @ 260 nm and 280 nm is < 0.1)
https://ecatalog.corning.com/life-sciences/b2c/US/en/Microplates/Assay-Microplates/96-Well-Microplates/Corning%C2%AE-96-well-UV-Transparent-Microplates/p/3635
As Dr Kanwar has Mentioned, ALISA Plates have a peak(s) of absorption
Approximately between 200-300 [nm] depending on plate's material.
Here is a sample absorption spectra of my samples. Blank well is somthing like these curves.
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