B-PER™ Direct Bacterial Protein Extraction Kit - FAQs

B-PER Reagent is designed to be used with, among other applications, 6xHis and GST fusion protein purification. When using B-PER Reagent with other IMAC or glutathione columns, use a dilution of at least one-to-one B-PER Reagent in the binding buffer recommended for that column. This will decrease the possibility of incompatibility of buffer components.

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

Bacteria grown in rich medium (high glucose) are difficult to lyse. Try reducing the glucose concentration. Bacteria and organisms lysed with B-PER Reagent include E. coli, Acetinobacter, Archaebacteria, S. aureus, H. pylori, Baculovirus, nematodes, and insect cells. B-PER Reagent can lyse some gram-positive bacteria, but tends to be more efficient with gram-negative bacteria. If lysis is inefficient for a particular bacterial strain, freeze cells before extraction. Add lysozyme for the most efficient extraction, however, for some over-expressed proteins, the addition of lysozyme is not required.

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

Many proteins spontaneously refold into their native, functional structures when the denaturing solubilization reagents are removed by dialysis. Other proteins, however, will fold into non-functional and even insoluble forms by this process. In such cases, specialized sets of buffer conditions must be tested to identify those that promote the highest possible yield of properly refolded protein. See our Pierce Protein Refolding Kit (Cat. No. 89867).

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

Add DNase I to the B-PER solution. If DNase was added, the enzyme may have low activity or be inactive. You can add Mg2+ ions to 2 mM final concentration, add more DNase I to the reagent, or purchase new enzyme. Alternatively, use B- PER Complete reagent, an all-in-one formulation that combines a lysis reagent with lysozyme and a universal nuclease to enable mild extraction from both gram-negative and gram-positive bacteria.

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

The temperature may be too low; warm the culture and other reagents to room temperature. The lysozyme may have low activity or be inactive. Add more lysozyme to the reagent or purchase new enzyme.

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

We recommend using B-PER Complete Bacterial Protein Extraction Reagent, an all-in-one formulation that combines a lysis reagent with lysozyme and a universal nuclease to enable mild extraction from both gram-negative and gram- positive bacteria. If the addition of lysozyme and DNase I might interfere with your downstream application, we recommend using other B-PER reagents. See details of the various B-PER reagents we offer.

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

Some overexpressed proteins can be insoluble, misfolded, or expressed in inclusion bodies. The best way to obtain soluble protein is to adjust the expression conditions; however, if this is not possible, the Inclusion Body Solubilization Reagent (Cat. No. 78115) can be used.

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

See details of the various B-PER reagents we offer (https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-purification-isolation/cell-lysis-fractionation/cell-lysis-total-protein-extraction/b-per-bacterial-cell-lysis-reagents.html).

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

We offer the B-PER Bacterial Protein Extraction Reagent for lysis of bacterial cells.

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