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HomeLysis & Protein ExtractionUse of BugBuster® and Lysonase™ Reagents for efficient protein extraction from Gram-positive bacteria

Use of BugBuster® and Lysonase™ Reagents for efficient protein extraction from Gram-positive bacteria

BugBuster® Protein Extraction Reagent has become a valuable tool for extraction of proteins from E. coli without the need for mechanical disruption. It is also known that supplementing BugBuster® (as well as its derivative, PopCulture™ Reagent) with a small amount of lysozyme enhances the efficiency of extraction from E. coli, especially for larger proteins1. Furthermore, the addition of Benzonase® Nuclease to the extraction mixture degrades the liberated nucleic acids, reducing viscosity and potential interference of DNA and/or RNA with purification procedures2. With this issue of inNovations, we introduce Lysonase™ Bioprocessing Reagent, which is an optimized blend of rLysozyme™ Solution and Benzonase® Nuclease that can be used alone or in conjunction with BugBuster® or PopCulture™ Reagent for any of their applications.

Figure 1 shows that the combination of BugBuster® and Lysonase™ greatly enhanced the release of soluble proteins from two different species of Gram-positive bacteria. Extraction is accomplished by resuspending cells in BugBuster® containing 1/125 volume of Lysonase™ followed by a 20-minute incubation at room temperature. Insoluble material is simply removed by centrifugation and the supernatant can be used directly for purification or activity assays.

Protein Extraction Gram Positive Bacteria

Figure 1. Combination of BugBuster® and Lysonase™ greatly enhanced the release of soluble proteins from two different species of Gram-positive bacteria. Extraction is accomplished by resuspending cells in BugBuster® containing 1/125 volume of Lysonase™ followed by a 20-minute incubation at room temperature. Insoluble material is simply removed by centrifugation and the supernatant can be used directly for purification or activity assays.

Materials

Product NumberProduct NameProduct DescriptionPricing
BM-100Auto2D® 2-D Electrophoresis Deviceautomates 2D gel electrophoresis
BM-1EAuto2D® Electrode ChipAuto2D® Electrophoresis Device
BM-1SAuto2D® Solution ChipAuto2D® 2-D Electrophoresis Device
BM-1EPAuto2D® Electrode Chip PlusAuto2D® Electrophoresis Device
BM-113010Auto2D® IEF Chip pH3-10precast gel, used for first-dimension separations on the Auto2D® 2-D Electrophoresis Device
BM-114055Auto2D® IEF Chip pH4-5.5for first-dimension separations on the Auto2D® 2-D Electrophoresis Device., precast gel
BM-116010Auto2D® IEF Chip pH6-10used for first-dimension separations on the Auto2D® 2-D Electrophoresis Device., precast gels
BM-117010Auto2D® IEF Chip pH7-10for first-dimension separations on the Auto2D® 2-D Electrophoresis Device
BM-12065Auto2D® PAGE Chip 6.5%for second-dimension separations on the Auto2D® 2-D Electrophoresis Device, precast gel
BM-12075Auto2D® PAGE Chip 7.5%precast gel, for second-dimension separations on the Auto2D® 2-D Electrophoresis Device.
BM-12100Auto2D® PAGE Chip 10.0%precast gel, for second-dimension separations on the Auto2D® 2-D Electrophoresis Device
BM-12125Auto2D® PAGE Chip 12.5%precast gel, for second-dimension separations on the Auto2D® 2-D Electrophoresis Device.
BM-1RYSJ1Auto2D® Glycine Type Reagent Kitreagent kit for the Auto2D® 2-D Electrophoresis Device
BM-1RYTJ1Auto2D® Tricine Type Reagent Kitreagent kit for the Auto2D® 2-D Electrophoresis Device, for better resolution on low molecular weight range

References

1.
Grabski A, Drott D, Handley M, Mehler M, Novy R. 2001. inNovations.(13):1-4.
2.
Welch ML, Anderson LL, Grabski WJ. 1999. EVALUATION AND MANAGEMENT OF NONMELANOMA SKIN CANCER. Dermatologic Clinics. 17(1):19-28. https://doi.org/10.1016/s0733-8635(05)70067-x
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