Protein Diluent Buffer

doi:10.1101/pdb.rec075234

Recipe

Protein Diluent Buffer

Buffer D (1×)	500 μL
50 μg/μL BSA (bovine serum albumin)	1 μL
14 m β-mercaptoethanol	1 μL

BSA is used here as a protein-stabilizing agent and β-mercaptoethanol as a reducing agent; 0.1–1 mm DTT can also be used. Reducing agents are particularly important for protein with cysteines within the DNA-binding domain (i.e., zinc fingers or the basic region of many bZIP proteins). Anti-aggregation agents such as 0.01%–0.1% NP-40 or Triton X-100 can also prove useful. Prepare this buffer in small quantities and store in aliquots in a freezer. Long-term storage of the solution on ice or in the refrigerator is not recommended.