B-PER™ Bacterial Protein Extraction Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Bacterial pellets are recovered after centrifugation (10,000rpm, 10 min), washed twice in PBS, and resuspended in 100 µl of B-PER II bacterial protein extraction reagent following the manufacturer's protocol.	- Culture supernatants are precipitated in 60% ammonium sulfate or trichloroacetic acid at 4°C overnight. After centrifugation, pellets are washed twice with 5ml cold acetone, dried, and resuspended in 200 µl of 2× Laemmli buffer.

Protocol tips
- Bacterial pellets are recovered after centrifugation (10,000rpm, 10 min), washed twice in PBS, and resuspended in 100 µl of B-PER II bacterial protein extraction reagent following the manufacturer's protocol.
Downstream tips
- Culture supernatants are precipitated in 60% ammonium sulfate or trichloroacetic acid at 4°C overnight. After centrifugation, pellets are washed twice with 5ml cold acetone, dried, and resuspended in 200 µl of 2× Laemmli buffer.

Publication protocol

L. monocytogenes strains were cultured at 37°C in 10 ml BHI or BHI supplemented with 0.2% charcoal (Merck). Bacterial pellets were recovered after centrifugation at 10,000 rpm for 10 min, washed twice in PBS, and resuspended in 100 µl of B-PER II bacterial protein extraction reagent (Pierce) to isolate total bacterial proteins. Culture supernatants were precipitated in 20, 40, 60, and 80% ammonium sulfate or trichloroacetic acid at 4°C overnight. After centrifugation, pellets were washed twice with 5 ml cold acetone, dried, and resuspended in 200 µl of 2× Laemmli buffer.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for B-PER™ Bacterial Protein Extraction Reagent below.

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