Overview: Lysis Buffers
Lysis buffers are used to break open cell walls and membranes. This enables the release of intracellular materials such as DNA, RNA, protein, lipids or organelles from a cell for later use in a wide array of biological experiments [1]. The selection of appropriate lysis buffers plays an important role in the preparation of cell lysates, the quality samples to be used for downstream processes. Such processes include protein purification for studying protein function and structure, enzymatic assays, immunoprecipitation, protein-protein interaction, SDS-PAGE, Western blotting, and ELISA.
Lysis buffers generally vary in composition and are specifically designed to lyse specific cellular compartments that contain analytes of interest. These buffers contain different chemicals specialized for degrading certain membranes. The selection of an ideal Lysis buffer for a specific biological experiment is dependent on the cell type being analyzed, the basic structure of the cell, and the location of the analyte of interest within the cell [2]. These buffers typically include a buffering component, an inorganic salt, a variable detergent such as a non-ionic detergent, an ionic detergents, or zwitterionic detergent, along with protease and phosphatase inhibitors, and chelators. With variable mixtures, Lysis buffers can be specially formulated for certain procedures.
Image 1: The mechanism of cellular lysis. Using a detergent within a lysis buffer solution, lysates may be extracted from specific cellular compartments for later analysis.
Common Lysis Buffers
Several Lysis buffers can be formulated to obtain high yields and purities of released proteins dependent on the location of the desired protein. The following Lysis buffer types that can be used for each cellular location as described below.
Lysis Buffer |
Description |
|---|---|
| RIPA (Radioimmunoprecipitation) Lysis Buffer | Causes rapid cell lysis and solubilization of proteins from various cellular compartments, including the nucleus and mitochondria. Effectively enables extraction of cytoplasmic, nuclear, mitochondrial, and membrane proteins. Because of its strong lysis properties, RIPA buffers may result in solubilization of non-specific proteins as well as disruption of protein-protein interactions, which may interfere with downstream applications. |
| NP-40 Lysis Buffer | Maintains the native form of proteins, suitable for the extraction of cytoplasmic or membrane-bound proteins or for the preparation of whole cell extracts. Moderate strength retains protein functions with minimal disruption. Used in immunoprecipitation, SDS-PAGE, Western blotting, and for studying protein-protein interactions. |
| Tris-Triton Lysis Buffer | Commonly used for extracting membrane- or cytoskeleton bound proteins. |
| NETN Buffer | Mild lysis buffer for extracting nuclear proteins, separating nuclear and cytoplasmic fractions. Mild lysis buffer for extracting nuclear proteins, separating nuclear and cytoplasmic fractions. |
| Tris-HCl Buffer | Mild strength lysis buffer suitable for solubilizing cytoplasmic proteins. |
| Triton X-100 Lysis Buffer | Moderate strength lysis buffer generally used to lyse cells for extracting proteins, especially membrane proteins under mild non-denaturing conditions. |
| CHAPS Lysis Buffer | Chaps is a non-denaturing and zwitterionic detergent used in the lysis buffer for the solubilization and purification of membrane proteins. A CHAPS lysis buffer exhibits strong cell lysis and protein extraction capacity while preserving protein conformation and protein-protein interaction for co-immunoprecipitation (Co-IP) analysis. |
| HEPES Lysis Buffer (with CHAPS) | Strong lysis buffer containing non-denaturing zwitterionic detergent CHAPS is ideal for general cell-tissue lysis, molecular fractionation, and solubilization of membrane proteins. Strong protein extraction capacity while preserving protein conformation and thus ideal for co-immunoprecipitation and analysis for protein-protein interaction. |
| HEPES Lysis Buffer (with NP-40) | Moderate strength lysis buffer resulting in extraction of proteins with minimal disruption and retention of protein functions intact. Suitable for protein analysis assays such as ELISA, SDS PAGE and Western blotting. |
| HEPES Lysis Buffer (with Triton X-100) | Moderate strength lysis buffer widely used to lyse cells for extracting proteins, especially membrane proteins under mild conditions. |
Analyte Location and Lysis Buffer Use
The most used lysis buffers in biochemical and molecular biological experiments are NP-40 and RIPA lysis buffers, which are suitable for obtaining whole-cell extracts. Since RIPA is a strong lysis buffer, it can lyse almost every cellular compartment. NP-40 lysis buffer is applicable to a variety of applications as well.
Location of Protein |
Lysis Buffer |
|---|---|
| Whole Cell | NP-40 or RIPA |
| Membrane Bound | NP-40 or RIPA |
| Nuclear | RIPA |
| Mitochondrial | RIPA |
| Cytoplasmic (soluble) Tris-HCl or RIPA | Tris-HCl or RIPA |
| Cytoplasmic (cytoskeletal bound) | Tris-Triton |
Lysis Buffers at Boston BioProducts
Every Lysis Buffer is unique to the cell type used and the experimental application. Select the appropriate Lysis Buffers from the catalog or design your optimal formulation with custom manufacturing options at Boston BioProducts.
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References:
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- Islam, M.S., Aryasomayajula, A and Selvaganapathy, P.R., (2017). A Review on Macroscale and Microscale Cell Lysis Methods. Micromachines, 8(3), 83; https://doi.org/10.3390/mi8030083.
- Peach, M., Marsh, N., & MacPhee, D. J. (2012). Protein solubilization: attend to the choice of lysis buffer. In Methods in molecular biology (Vol. 869, pp. 35-42). Protein Solubilization: Attend to the Choice of Lysis Buffer | SpringerLink