Bacterial PE LB™ has been developed for the extraction of soluble proteins and inclusion bodies from bacterial cells. It is a proprietary improvement on the lysozyme based lysis, which allows extraction of soluble proteins and concurrent removal of nucleic acids (DNA & RNA) released during cell lysis.Bacterial PE LB™ lysis eliminates viscosity build-up, allowing effective clarification with lower centrifugal force.
This kit is provided with an optional protocol for the formation of spheroplast and removal of lytic enzyme (Lysozyme) prior to lysis and extraction of the bacterial proteins. Bacterial PE LB™ is based on organic buffering agents and utilizes a mild non-ionic detergent and a proprietary combination of various salts and agents to enhance extraction and stability of proteins.Depending on the application, additional agents such as reducing agents, chelating agent, and protease inhibitors cocktail may be added into Bacterial PE LB™. This reagent has been tested for use with several widely used bacteria including E. coli strains.
Bacterial PE LB™ is compatible with most downstream applications including running various chromatography, gel electrophoresis applications, and protein folding procedures. Bacterial PE LB™ is also compatible for protein estimation with NI™ protein assay (Non-Interfering Protein Assay)™ .
For bacterial lysis, the kits are suitable for extracting soluble proteins from approximately 30g wet cell pellets for every 100ml Bacterial PE LB™. When the kits are used for extracting soluble proteins from spheroplasts, they are suitable for approximately 9g wet cell pellets for every 25ml Bacterial Suspension Buffer supplied.
Bacterial PE LB™ Formats
Complete Kits (Cat. # 786-176, 786-187, 786-188)
The complete kits feature Bacterial PE LB™, bacterial suspension buffer and PE LB™ Lysozyme .The PE LB™ Lysozyme is a proprietary mix of lysozyme, to break open cells, and DNase and RNase, to remove nucleic acids.The enzyme mix is ready-to-use and at concentrations for optimal enzymatic activity.
Bacterial PE LB™ buffer only (Cat. # 786-177, 786-185, 786-186)
The buffer only option allows researchers to substitute the Bacterial PE LB™ buffer into their existing protocols for improved lysis or for use in further downstream applications.
Bacterial PE LB™ [2X] (Cat. # 786-189)
Bacterial PE LB™[2X] is a modified formulation of Bacterial PE LB™ that allows half the volume of lysis buffer to be used while maintaining lysis rates. It is used for extraction of proteins that are expressed at low levels and can also be used when high concentrations of extracted proteins are desired.
Bacterial PE LB™ in Phosphate Buffer (Cat. # 786-191)
Bacterial PE LB™ in Phosphate buffer is a variation of Bacterial PE LB™ buffers only in phosphate buffer. It has the same efficiency as Bacterial PE LB, however the resulting lysates can be used for protein labeling and coupling reactions that use primary amines.
Features
- Multiple formats available: Complete kits with lytic enzymes; Buffer only; 2X buffer format; Phosphate buffer format
- Eliminates laborious mechanical lysis
- Eliminates removal of nucleic acids
- Eliminates viscosity build-up
Applications
- Suitable for preparation of spheroplast, lysis and extraction of proteins from bacterial cells and isolation of inclusion bodies
- Lysis and extraction of proteins from bacterial cells
- Isolation of biologically active bacterial and recombinant proteins