I was just wondering why besides the 280 nm wavelength there is an option for 254 nm in the Akta Prime plus protein purification system. From a google search I only got to know that the absorbance of proteins at 254 is half the absorbance at 280 nm.
Well the 280 nm filter in the Akta prime plus in our lab is not working properly, as we were getting flat absorbance even with standard proteins. So now we are using the 254 nm filter till the time 280 nm filter is replaced and we are getting good sharp peaks.
It is a detector i believe and consists of diode that detects proteins at 280 nm wavelength as protein shows absorption at 280 nm due to aromatic amino acids and due to double bonds where ever present. I dont know what is happening at 254 nm wavelength, Allah knows better may be some new thing has got introduced in it.... how about the activity of that purified protein?
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