Dear Sir/Madam,
I am having a solution containing less than 10 kDa peptides. How to quantify the peptide content in terms of concentration of peptides per mL by using Spectrophotometer? what is the wavelength to be used to measure peptide using a spectrophotometer? Kindly suggest a suitable method.
Hi,
You could consider using 214nm. Does any of your peptides contain trypthophans or other aromatic amino acids? If so, 279/280nm and/or 295nm could be an option too.
Be sure to also always check the absorbance at the chosen wavelength of the buffer you're using (without) peptides.
Hope this helps!
Sir,
Thank you for your kind suggestion.
I need more clarification.
The protein was cleaved at amino acids phenylalanine, tryptophan and tyrosine by enzymatic hydrolysis. Therefore, the peptides obtained had an amino acid sequence which contains phe, try, and tyr at the end of the chain.
In this case what wavelength to be used to determine peptide content?
The answer to your question depends on you standard and/or sample matrix. It could be Lowry, Biuret, or the Bradford color method.
Sir,
AS per the suggestions I received we can measure the peptide content directly taking the peptide solution in the cuvette and measuring at 205/210/214 or even at 280 nm. But I want to confirm which wavelength to be used finally.
Any suggestions plz.
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