Should I add b-mercaptoethanol to the lysis buffer?
Reducing agents can reduce the resin matrix and adversely affect binding of the His-tagged protein to the spin column. Its inclusion depends upon whether the His-tagged protein elutes with contaminants as -mercaptoethanol can reduce all disulphide bonds formed between the contaminating proteins and the target protein. Concentration less than or equal to 10 mM -mercaptoethanol can be used with Proteus IMAC resin spin columns. Do not use strong reducing agents such as DTT or DTE as these tend to reduce the metal ion, which will lower the binding efficiency of the IMAC column.
Related Questions
- Can I use other types of cell lysis and/or extraction buffer instead of the Extraction buffer provided in the Array Assay Kit?
- Can the Bioruptor® Sonicator be used to lyse cells instead of using the lysis buffer which is included in the MeDIP kit?
- Is it necessary to add the protease inhibitor to the lysis buffer?
