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

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Cell cultures centrifuged at 14,000 rpm for 5 min. - Supernatants were passed through an 0.2-μm Acrodisc syringe filter.	- Cell pellets are resuspended in 100 μl of B-PER protein extraction reagent and incubated for 10 min at room temperature. After, they are centrifuged at 14,000 rpm for 10 min, and supernatants are recovered. - If there are insoluble precipitates (from membrane fractions) they can be resuspended with 50 μl of B-PER protein extraction reagent.

Upstream tips
- Cell cultures centrifuged at 14,000 rpm for 5 min. - Supernatants were passed through an 0.2-μm Acrodisc syringe filter.
Protocol tips
- Cell pellets are resuspended in 100 μl of B-PER protein extraction reagent and incubated for 10 min at room temperature. After, they are centrifuged at 14,000 rpm for 10 min, and supernatants are recovered. - If there are insoluble precipitates (from membrane fractions) they can be resuspended with 50 μl of B-PER protein extraction reagent.

Publication protocol

One-milliliter aliquots of aerobic or anaerobic cultures of PAO1 grown to similar final cell densities (optical density at 600 nm [OD600] of ∼3.0) were centrifuged at 14,000 rpm for 5 min. Supernatants were passed through an 0.2-μm Acrodisc syringe filter (Pall Life Science Inc., Ann Arbor, MI) and saved as culture supernatants. Cell pellets were resuspended in 100 μl of B-PER protein extraction reagent (Thermo Fisher Scientific Inc., Rockford, IL) and incubated for 10 min at room temperature. Lysed cells were centrifuged at 14,000 rpm for 10 min, and supernatants were recovered (cell extract fractions). Insoluble precipitates were then resuspended with 50 μl of the same B-PER protein extraction reagent (membrane fractions).

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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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