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

Protein isolation Bacteria - Escherichia coli

Experiment
Protein isolation Bacteria - Escherichia coli
Product
B-PER™ II Bacterial Protein Extraction Reagent (2X) from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Upstream tips
- Capable of extracting bacterial microcompartments (BMCs).
Protocol tips
- The manufacturer protocol was followed.
Downstream tips
- Compatible with most downstream workflows such as electrophoresis, affinity purification, immunoprecipitation, protein interaction analysis, crosslinking and protein labeling.

Publication protocol

In order to characterize and compare the structural and biochemical properties of wtBMCs, rBMCs and eBMCs, a number of purification methods to isolate intact BMCs were investigated involving the use of mild detergent lysis procedures, employing either a bacterial specific lysis reagent (B-PER™) or a yeast specific lysis reagent (Y-PER™). The bacterial protein extraction (B-PER™) method is based on lysing cells with B-PER™ and entails consecutive centrifugation steps to separate cell debris from Pdu metabolosomes. This procedure was found to work best for the isolation of complete BMCs (wtBMCs and rBMCs). The yeast protein extraction reagent (Y-PER™) method is based on lysis of bacterial cells and subsequent salt precipitation and was found to be effective for the purification of eBMCs. The purification and isolation of the various BMCs was followed by SDS-PAGE.

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Discussion

Discussion

5 years ago

Author: Hollie Fowler United Kingdom

How can I deal with my pellet being too viscous?

I am using 8M Urea to lyse my cells but after centrifugation my pellet is very viscous. This causes me trouble when I try to collect the supernatant. Is there any way to avoid it and do you think it will have an effect on the concentration of my protein?

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Papers

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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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