What other buffers are used to quench the label reaction in the HDX workflow, besides what is in the Enzymate BEH Pepsin care and use manual? - WKB117316

ENVIRONMENT

• HDX workflow
• Enzymate BEH Pepsin column

ANSWER

Buffers that have been used include the following: 4M guanidine HCL and 0.5M TCEP, 3M guanidine HCL and 0.4M TCEP, 200mM phosphate buffer with 0.5M TCEP and 4M guanidine HCL pH 2.3,  and 33mM HCL in water pH 2.5.

ADDITIONAL INFORMATION

Change of quenching buffer may necessitate optimization of digestion timing, flow rate, and temperature. In addition, some buffers will produce more background noise, and care must be taken to reduce precipitation in the LC/MS system.

For additional information, see HDX: How to desalt from TCEP contamination and HDX: How to filter solvents and buffers.

See also the Enzymate BEH Pepsin Columns Care and Use Manual.