Description: NZY Bacterial Cell Lysis Buffer is an innovative product developed by NZYTech for the gentle disruption of Escherichia coli cell wall, generating a homogeneous cell-free extract. It provides a rapid and cost-effective alternative to harsh chemicals or mechanical procedures, such as French Press or sonication, to release recombinant and native proteins without denaturation. This extraction reagent is a Tris-buffered formulation (pH 7.5) with lysozyme and DNase I provided separately. The addition of these enzymes allows a most efficient extraction.
In E. coli, protein extraction efficiencies are strain-dependent. NZY Bacterial Cell Lysis Buffer can be used to disrupt most common bacterial host strains and it is especially efficient with BL21 strain and its derivatives. The use of pLysS or pLysE hosts enhances protein extraction efficacies.
– Ready-to-use formulation
– Outstanding yields of extracted proteins
– Non-denaturing protein extraction
– DNase I and Lysozyme provided separately
– Disrupt E. coli cell walls without mechanical procedures
– Ideal for high-throughput (HTP) methods
Easy and effective protein extraction from cell-free extracts
NZY Bacterial Cell Lysis Buffer is as efficient as standard mechanical lysis (ultra-sonication) to disrupt E. coli cells while retaining the biological activity of the recombinant proteins. Detergent lysis is also more amenable in HTP approaches, when you have to disrupt multiple recombinant cell strains simultaneously.
5-mL samples of twenty two 40-mL cultures of BL21(DE3) cells expressing different Ruminococcus flavefaciens proteins were harvested by centrifugation and pellets were either ressuspended in 1 mL of 50 mM NaHEPES buffer pH 7.5, followed by sonication or lysed in 1 mL of NZY Bacterial Cell Lysis Buffer, supplemented with 0.1 mg/mL lysosyme and 0.004 mg/mL DNase I (~15 min at room temperature). Recombinant proteins extracted using these two strategies were purified through IMAC and separated by SDS-PAGE. Protein concentration was determined using a NanoVue spectrophotometer. Overall, the cell-free extracts from the biochemical lysis showed higher to similar levels of target protein extracted when compared with sonication.
Storage conditions: Store at room temperature. Lysozyme and DNase I should be stored at -20 ºC
Shipping conditions: Shipped at 4 ºC to room temperature
– NZY Bacterial Cell Lysis Buffer
– Lysozyme (at 50 mg/mL)
– DNase I (at 2 mg/mL)