Is it possible to identify and quantitate the disulfide bonds in cysteine-containing residues in proteins using the three-step protocol of the ProteinWorks Auto-eXpress Low 3 Digest Kit (176004077)? - WKB77536

ENVIRONMENT

• ProteinWorks Auto-eXpress Low 3 Digest Kit
• Xevo TQ-S

ANSWER

The three-step protocol will not impact the disulfide bonds in the cysteine-containing residues. The disulfide bonds can be quantitated with the use of mass spectrometry. 

ADDITIONAL INFORMATION

It will be very difficult to quantitate the disulfide bonds in the samples without mass spectrometry. If using only a UV, there will no doubt be variations in retention time. In addition, because this is mainly a mass spectrometry application, the amount of digestion buffer, trypsin solution, digestion inactivation reagent, and other agents may need to be greatly increased to quantitate in UV applications. Waters does not have data that the disulfide bonds in some of the peptides will have good sensitivity in the UV range.The ProteinWorks Auto-eXpress Low 3 Digest Kit (176004077) is an automation kit with which the quantity of each reagent/standard is optimized for the MS application. If done without mass spectrometry, reagent volumes, incubation times, and incubation temperature must be optimized.