Lysozyme Solution (50 mg/mL)

The Thermo Scientific BCA Protein Assay Kit (Cat. No. 23225, 23227) works very well to detect proteins extracted with B-PER Reagent. The Coomassie Plus Protein Assay (Cat. No. 23236) can be used with B-PER Reagent Protein extractions, however when using B-PER with PBS or B-PER II reagents, dilute the protein preparation two-to-four fold before assaying with Coomassie Plus or Prepare sample with Compat-Able Protein Assay Preparation Reagent Set (Cat. No. 23215). Finally, Thermo Scientific 660 nm Assay Kit (Cat. No. 22662) can also be used if protein extract is diluted two fold.

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

Add 1-2 µL of 50 mg/mL Lysozyme Solution (Cat. No. 90082) to 200 µL of the EasyPep lysis buffer provided in the kit and proceed with the sample prep. Add 200 µL of the prepared lysis solution to 5 X 10E8 bacterial cells (E. coli cells) (OD600 equal to 1, corresponding to 1 X 10E8 CFUs). Lyse by pipetting the sample repeatedly and centrifuge at 16,000 x g for 5 mins. Alternatively, E. coli cells can be lysed in lysis buffer using bead beating or sonication instead of lysozyme. After lysis, dilute samples to 1 mg/mL using lysis buffer, proceed with the EasyPep protocol for reduction, alkylation, digestion, and clean up as described in the product manual.

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.

The B-PER Reagent solution contains a proprietary, mild, non-ionic detergent in 20 mM Tris-HCl, pH 7.5. It effectively disrupts cells and solubilizes native or recombinant proteins without denaturation. The reagent creates holes in the cell membrane that will leak out cytosolic proteins. The sample may become very viscous when the bacterial chromosome is released. We recommend adding DNAse I (Cat. No. 90083) to the reagent to reduce viscosity. For better lysis efficiency and if there are inclusion bodies, we recommend adding Lysozyme (Cat. No. 90082) to the reagent. Alternatively, you may purchase the B-PER Bacterial Protein Extraction Reagent with Enzymes Kit (Cat. No. 90078 or 90079) that includes the B-PER Bacterial Protein Extraction Reagent, DNase I, and Lysozyme.

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