ProBond™ Purification System

Western blot analysis is typically used to detect the expressed protein. We sell several antibodies against various epitopes, such as Xpress, HisG, V5, or C-terminal 6xHis. Additionally, His-tagged proteins can be purified using our ProBond Purification System via affinity purification.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

There are a few suggestions provided from our R&D scientists:

(1) Use stepwise dialysis against successively lower concentrations of urea buffers to slow the refolding.

(2) Add glycerol to the folding buffer; usually between 10 to 50%, but the amount must be determined empirically.

(3) After the denaturing washes, do several washes under native conditions and then elute under native conditions.

Note: you may also try to rescue a precipitated protein by adding denaturing buffer and either trying the glycerol dialysis or rebinding to the column.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

In one experiment, 35 µg CAT/mg total protein or 68.2 mg CAT/liter of culture was obtained. 50 ml of cell extract was loaded onto a ProBond column and 2 mg of CAT was recovered. The eluted protein appeared reasonably pure on a gel.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

The enzyme could be denatured. Try buffer exchange or dialysis before digestion with EKMax Enterokinase.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

ProBond and Ni-NTA beads can be used in FPLC columns. However, the beads can only withstand low pressure (~43.5 psi max).

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.