I think so, you just have to adjust the volumes off each reagent. Nevertheless, be aware that depending on your microplate reader, some wavelengths have too much noise ex; 220 nm to do the hydrogen peroxide assay.

Yes. Besides the adjustment of the volumes of each reagent, you will need to adjust the protein of the sample (between 0,5 and 1 mg/mL will be fine)

Hi Irmak

I have made this adaptation to microplates since I work with oocytes and the samples are too small for the spectrophotometer. I have measured proteins by Bradford, SOD, TBA-RS, and GSH. They work well but I think it is important that you have a reference curve to adjust the reagent and sample volumes. If you can do this, this will give you more confidence in your results. If you have a reference curve of known concentrations, and you know what values ​​it gives you in the spectrophotometer, you can compare with the values ​​obtained in the microplate reader. I hope this helps you and for any questions you can write me.

Regards

Respected Bilgiseven,

I fully agree with the above. It depends on the type of assay and its sensitivity , specificity, and wavelength. A colotimetric

vs U V Spectrophotometric assay. Colorimetric assays are less sensitive, hence you need to have high (0.250 microgram and above) conc and a Chromogen at 400-700 nm spectral wavelengh, whereas a U.V Spectrophotometer (260-280 nm wavelength) can be used at very low conc (ng ranges) for even colorless sample if your sample is very costly and you want to use it for some other estimations also like DNA, RNA, Protein, Enzyme, hormone, cytokine, or chemokine , antibody, antigen estimations etc. Fluorescent Spectrometric assays are still more sensitive. If you have FRET or SPR it will add more luster to your experimental efforts. But these are costly and require little experties.

I hope it is informative. Good luck with your research.

Respectfully,

Dean Sharma

Every procedure is strictly according to manufacturers standard operating procedure . Spectrophotometric procedure is different from that of microplate reader. Most microplate reader uses ELISA method

Firstly by adjusting the volume, you can precisely use it; by adjusting both your sample and the standard, blank or whatever it is you will use to the same volume. After choose the absorbance mode on the microplate reader, the working principle is the same with a UV-vis-spectrophotometer. Besides, instead of spending 1 mL sample for each reading, you can make 3 replicas from 100x3= 300 ul.

But if you working with a wavelength from the UV region of the light spectrum (for example, CAT assay may need the absorbition of H2O2, 230 nm), unfortunately you cannot read them with those plastic plates.

Hello, first you must know the aim of the test (quantitative or qualitative) then to read some papers about the incertitude and limitation of the second technique if it seems that it is okay you should read the comparison between the two techniques and to be confident that there is no significant difference between ordinary spectrometric method and the microplate