B-PER™ II Bacterial Protein Extraction Reagent (2X)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- B-PER treatment did not solubilize the cells, instead it permeabilized the cells.	- Cells are resuspended in one-tenth of the original volume in the B-PER Bacterial Protein Extraction Reagent (Thermo Scientific, U.S.) containing EDTA-free protease inhibitor (Roche, Switzerland) and are shaken gently for 10 min in ice before centrifugation at 5000g for 10 min at 4 °C.

Upstream tips
- B-PER treatment did not solubilize the cells, instead it permeabilized the cells.
Protocol tips
- Cells are resuspended in one-tenth of the original volume in the B-PER Bacterial Protein Extraction Reagent (Thermo Scientific, U.S.) containing EDTA-free protease inhibitor (Roche, Switzerland) and are shaken gently for 10 min in ice before centrifugation at 5000g for 10 min at 4 °C.

Publication protocol

Cultures with an optical density value at 730 nm cm−1 between 1 and 2, or cultures that have been treated with chemicals, were harvested by centrifugation for 10 min at 5000g at 4 °C. The cells were resuspended in one-tenth of the original volume in the B-PER Bacterial Protein Extraction Reagent (Thermo Scientific, U.S.) containing cOmplete ethylenediaminetetraacetic acid (EDTA) free protease inhibitor (Roche, Switzerland) (one tablet in 25 ml of the B-PER reagent), and were shaken gently for 10 min in ice before centrifugation at 5000g for 10 min at 4 °C. The pellets were subsequently resuspended in one-fourth the original culture volume of 20 mM Tris–HCl buffer, pH 7.5. The control samples were subjected to the same protocol, except that the B-PER reagent was replaced with the 20 mM Tris–HCl buffer, pH 7.5. The supernatants were discarded unless otherwise stated, and the pellets were resuspended in the buffer, which were used directly for enzyme activity assays and for determination of the chlorophyll a (Chl a) content.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for B-PER™ II Bacterial Protein Extraction Reagent (2X) below.

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