QIAGEN: Qproteome™ Bacterial Protein Preparation Handbook.

Qiagen Iranian Bio Data Center

In order to be able to efficiently purify and analyze proteins, they must first be released from their host cell in a soluble form. Therefore, the first step in protein purification and analysis is cell lysis. Bacteria possess a rigid cell wall composed of peptidoglycans that must be disrupted to access cellular proteins. This has traditionally been accomplished by mechanical means, for example by subjecting cells to shear forces in a French press or by sonication. However, these methods are time-consuming and labor-intensive, and often lysis is incomplete. The Qproteome Bacterial Protein Prep Kit provides gentle but efficient detergent-based lysis of bacterial cells to deliver a soluble protein fraction suitable for any downstream application. Cell pellets are incubated with Native Lysis Buffer that is supplemented with lysozyme and Benzonase®, enzymes which help disrupt cell walls and digest nucleic acids respectively. No specialized equipment is required. Cells are simply incubated with lysis buffer and centrifuged to separate cell debris and insoluble protein (e.g., inclusion bodies). The supernatant contains the soluble protein fraction, which can be further purified or directly analyzed, for example by SDS-PAGE

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