Search Thermo Fisher Scientific
Cell Lysis Buffers |
Cell lysis is a fundamental step in biological research that involves breaking open cell membranes to release cellular contents, allowing for the extraction and study of intracellular components such as proteins, DNA, RNA, and organelles. Cell lysis buffers are central to successful cell lysis. The careful selection of the most suitable lysis buffer is crucial for obtaining high-quality samples and accurate results in a wide range of applications. Cell lysis buffers are specially formulated to effectively disrupt cell membranes while maintaining the stability and activity of intracellular components. They typically contain detergents, salts, chelating agents, and protease inhibitors tailored to specific cell types and compatible with downstream applications such as western blot, ELISA, protein purification, immunoprecipitation, mass spectrometry, and gene expression analysis.
Thermo Fisher Scientific offers a range of lysis buffers specifically designed to address the distinct characteristics of mammalian, bacterial, plant, yeast, and insect samples. Mammalian cells commonly use RIPA buffer as it contains detergents and salts that can efficiently lyse cells and offer the flexibility to add protease inhibitors for maintaining the integrity of extracted proteins. Bacterial cells require B-PER or BugBuster which contains enzymes or detergents that effectively break down the bacterial cell wall. Specialized buffers are available for the unique characteristics of plant, yeast, and insect samples. Cell lysis buffers have been optimized to delicately disrupt cell membranes and extract cellular contents with precision and efficiency, while maintaining their integrity and functional activity. By understanding the types of lysis buffers available for different sample types, researchers can choose the lysis buffer that is most suitable for their experimental needs to help ensure reliable protein extraction and accurate downstream analysis. Choose from our comprehensive range of cell lysis buffers based on the specific needs of your experiment.
On this page:
Key features of cell lysis buffers
- Specificity—designed for specific sample types including mammalian cells, bacteria, plant, yeast, and insect cells helping ensure optimal performance
- Efficiency—offer efficient and rapid cell membrane disruption helping ensure maximum recovery of intracellular content while minimizing sample loss
- Preservation—preserve the integrity and functionality of proteins, DNA, RNA, and organelles preventing degradation and maintaining the functional activity
- Compatibility—compatible with a variety of downstream applications, while safeguarding the quality of samples for downstream analysis
Choose the best lysis buffer for your experimental needs
Cell lysis buffers for mammalian cells often contain detergents, salts, and other solutions to disrupt the lipid bilayer of the cell membrane and extract proteins, DNA, RNA, and other cellular components. These lysis buffers include the option to add protease or phosphatase inhibitors to aid in stabilizing the extracted sample.
Cell lysis buffers for mammalian culture cells
| RIPA Lysis Buffer | IP Lysis Buffer | M-PER Mammalian Protein Extraction Reagent | NP-40 Cell Lysis Buffer | Cell Lysis Buffer | |
| When to use | Extraction of cytoplasmic and nuclear protein in denaturing conditions | Formulated specially for pull-down and immunoprecipitation assays | Mild, non-denaturing and efficient lysis for cytoplasmic and nuclear protein extraction | Recommended for extraction of cytoplasmic proteins | Need a harsher buffer than NP-40 or when cytoplasmic and nuclear protein extraction is needed |
| Composition | 25 mM Tris-HCl, pH 7.6 150 mM NaCl 1% NP-40 1% sodium deoxycholate 0.1% SDS | A modified RIPA buffer formulation without SDS | Non-denaturing detergent in 25 mM bicine buffer (pH 7.6) | 50 mM Tris, pH 7.4 250 mM NaCl 5 mM EDTA 50 mM NaF 1 mM Na3VO4 1% NP-40 0.02% NaN3 | 10 mM Tris, pH 7.4 100 mM NaCl 1 mM EDTA 1 mM EGTA 1 mM NaF 20 mM Na4P2O7 2 mM Na3VO4 1% Triton X-100 10% glycerol 0.1% SDS 0.5% deoxycholate |
| Recommended supplemental protease/phosphatase inhibitor(s) | Halt Protease Inhibitor Cocktail | ||||
| Compatible protein assays | BCA assays, Detergent Compatible Bradford | BCA assays, Detergent Compatible Bradford | BCA assays, Bradford assays | BCA assays, Detergent Compatible Bradford | Detergent Compatible Bradford |
| Downstream compability | SDS-PAGE, western blotting | Immunoprecipitations, pull-down assays | IP, protein electrophoresis, western blots, ELISA, EMSA, reporter assays, purification, activity assays, amine reactive labeling | ELISA, protein electrophoresis, western blotting | ELISA, protein electrophoresis, western blotting |
| Available size(s) | 100 mL 250 mL | 100 mL 250 mL | 25 mL 250 mL | 100 mL | 100 mL |
| User guide | User Guide: RIPA Lysis and Extraction Buffer | User Guide: Pierce IP Lysis Buffer | User Guide: M-PER Mammalian Protein Extraction Reagent | User Guide: NP-40 Cell Lysis Buffer | User Guide: Cell Lysis Buffer |
Cell lysis buffers for mammalian tissues
| T-PER Tissue Protein Extraction Reagent | N-PER Neuronal Protein Extraction Reagent | |
| When to use (compatible sample types) | Heart, liver, kidney, lung, spleen | Brain tissue and primary neurons |
| Composition | Non-denaturing detergent in 25 mM bicine, pH 7.6 150 mM NaCl | -- |
| Compatible protein assays | BCA assays, Bradford assays | BCA assays, Detergent Compatible Bradford |
| Downstream compatibility | IP, western blots, ELISA, EMSA, purification, kinase assays, activity assays, amine reactive labeling | IP, pulldowns, western blots, ELISA, enzyme assays, amine reactive labeling |
| Available sizes | 500 mL | 100 mL |
| User guide | User Guide: T-PER Tissue Protein Extraction Reagent | User Guide: N-PER Neuronal Protein Extraction Reagent |
Mammalian cell lysis reagents for cultured cells
RIPA lysis buffer
RIPA (radioimmunoprecipitation assay) is a popular cell lysis option that allows for the extraction of membrane, cytoplasmic, and nuclear proteins from cells or tissues. This lysis buffer is particularly effective because of its non-ionic and ionic detergents such as SDS and sodium deoxycholate. Alternatively, these components may decrease activity of some protein kinases or enzymes, thus it is advised to prepare a RIPA buffer that does not contain SDS and sodium deoxycholate. Protease or phosphatase inhibitors can be utilized with RIPA buffer to prevent proteolysis and preserve the phosphorylation of proteins.
IP lysis buffer
Thermo Scientific Pierce IP lysis buffer is a whole cell lysis buffer based on a modified RIPA buffer formulation without SDS. This moderate-strength lysis buffer effectively solubilizes cellular proteins but does not liberate genomic DNA or disrupt protein complexes like ordinary RIPA buffer. IP lysis buffer is an alternative cell lysis option to RIPA lysis buffer when a protein affinity purification experiment, such as immunoprecipitation (IP) or co-immunoprecipitation (CoIP), is the downstream application (Figure 1). This lysis buffer was formulated specially for pull-down and immunoprecipitation assays when it is especially important that the lysis reagent does not contain denaturants or components that might interfere with the antibody binding or protein interactions of interest. Protease or phosphatase inhibitors can be used to prevent proteolysis and maintain the phosphorylation of proteins.
Figure 1. Immunoprecipitation of cell lysates extracted using different lysis buffers. A549 cell lysates (200µg) were generated using different lysis buffers. EGFR and PP2A were immunoprecipitated using the Thermo Scientific Pierce Crosslink IP Kit and anti-EGFR antibody (Stressgen) or anti-PP2A antibody (Millipore). The immunoprecipitations were washed with the same lysis buffer used to lyse the cells. Lysates representing 5% of the total load (L) and elution (E) fractions were blotted with anti-EFGR or anti-PP2A antibody and using Thermo Scientific SuperSignal West Dura Substrate for detection.
M-PER mammalian protein extraction reagent
Thermo Scientific M-PER Mammalian Protein Extraction Reagent is an effective yet milder alternative to RIPA buffer. This reagent can extract soluble protein from primary, suspension, and adherent cell types (Figure 2). Lysis can be completed in only 5 minutes and directly added in culture plates, requiring little to no additional cellular disruption. M-PER can result in significantly more extracted protein compared with freeze/thaw and sonication methods (data not shown). Cell lysates obtained using M-PER are compatible with several downstream assays including immunoassays, enzyme assays, and various reporter assays. Additionally, M-PER contains dialyzable components without primary amines, making it suitable for luciferase, beta-galactosidase, and CAT assays unlike alternative protein extraction methods.
Click image to enlarge
Figure 2. Protein yield from various cell types using M-PER Mammalian Protein Extraction Reagent. Cells were harvested at 85% confluency, washed twice and collected in ice-cold PBS, and counted. For each cell type, 1 x 106 cells were pelleted by centrifugation at 2,000 x g for 5 minutes and lysed in 1 mL M-PER Reagent for 5 minutes. The cell lysates were clarified by centrifugation at 14,000 x g for 10 minutes, the supernatant was collected and the protein concentration (μg/million cells) was determined using the Pierce BCA Protein Assay.
Mammalian cell lysis reagents for tissues
Thermo Scientific T-PER Tissue Protein Extraction Reagent is designed for total protein extraction from mammalian tissue samples, including heart, liver, kidney, lung, and spleen (Figure 3). The T-PER reagent utilizes a non-denaturing detergent in 25 mM bicine, 150 mM NaCl (pH 7.6), and is used in conjunction with mechanical or manual homogenization. Furthermore, T-PER reagent was validated for protein extraction from fresh and frozen tissues.
Thermo Scientific N-PER Neuronal Protein Extraction Reagent is designed for total protein extraction from mammalian neuronal tissue samples or primary cultured neurons. N-PER lysis can be completed in less than 30 minutes and is used in conjunction with mechanical or manual homogenization. Protease or phosphatase inhibitors can be added to prevent proteolysis or protect against phosphatase activity normally present in brain tissue.
Click image to enlarge
Figure 3. Protein extraction from various tissue types using T-PER Tissue Protein Extraction reagent. Duplicate tissue samples were weighed, resuspended in 1:10 to 1:20 w/v T-PER reagent, and disrupted in a chilled Dounce or benchtop tissue homogenizer. The resulting lysates were centrifuged at 10,000 x g for 5 minutes and the supernatant was collected. The protein concentration of each lysate was determined using the Pierce BCA Protein Assay to determine protein yield per milligram of starting tissue.
Click image to enlarge
Figure 4. Protein extraction from mouse brain neuronal tissue using N-PER Neuronal Protein Extraction reagent. Homogenization of mouse brain neuronal tissue in Thermo Scientific N-PER Reagent resulted in greater overall protein yield than in our standard T-PER Tissue Protein Extraction Reagent and two varieties of phosphate-buffered saline (PBS).
B-PER Bacterial Protein Extraction Reagents gently lyse E. coli and other species of bacterial cells and effectively extract soluble native and recombinant proteins. Several different ready-to-use formulations of B-PER reagent are available for different lysis needs. Depending on downstream application, protease inhibitors, salts, reducing agents, denaturants, and chelating agents may be added.
| B-PER Complete Bacterial Protein Extraction Reagent | B-PER Bacterial Protein Extraction Reagent | B-PER II (2X) Bacterial Protein Extraction Reagent | B-PER (PBS) Bacterial Protein Extraction Reagent | |
| When to use (compatible sample types) | E. coli, B. subtilis | E. coli, B. subtilis | E. coli, B. subtilis | E. coli, B. subtilis |
| Features |
|
|
|
|
| For use with fresh and frozen cells? | Yes, add EDTA to gram-negative fresh cells | Yes | Yes | Yes |
| Composition | B-PER reagent formulated in Tris buffer with enzymes (lysozyme & universal nuclease) | B-PER reagent formulated in Tris buffer | 2X B-PER reagent formulated in in Tris buffer | B-PER reagent formulated in PBS |
| Nuclease and lysozyme in formulation? | Yes | No | No | No |
| Sample processing time | 15 min | 10–15 min | 10–15 min | 10–15 min |
| Mechanical disruption required? | No | No, but for gram-positive bacteria add enzymes for increased yield | No, but for gram-positive bacteria add enzymes for increased yield | No, but for gram-positive bacteria add enzymes for increased yield |
| Amount of sample processed | 100 g cell paste per 500 mL | 125 g cell paste per 500 mL | 125 g cell paste per 250 mL | 125 g cell paste per 500 mL |
| Compatible protein assays | BCA, Bradford assays | BCA, Bradford assays | BCA, Bradford assays | BCA, Bradford assays |
| Downstream compatibility | SDS-PAGE, western blotting, protein purification | SDS-PAGE, western blotting, protein purification (except for GST fusion proteins) | SDS-PAGE, western blotting, protein purification (except for GST fusion proteins) | SDS-PAGE, western blotting, protein purification (except for GST fusion proteins) |
| Available size(s) | 250 mL 500 mL | 165 mL 250 mL 500 mL | 250 mL | 500 mL |
| User guide | User Guide: B-PER Complete Bacterial Protein Extraction Reagent | User Guide: B-PER Bacterial Protein Extraction Reagent | ||
B-PER Bacterial Protein Extraction Reagents
B-PER reagents can be used for various Gram-negative and Gram-positive bacteria such as S. aureus, H. pylori, and E. coli strains BL21(D3), JM109, DH5a, and M15. The reagent is also suitable for certain Archaebacteria species and cultured insect cells. Extraction does not require expensive equipment or mechanical disruption and can be performed in 10 to 15 minutes. B-PER reagent removes soluble protein from inclusion bodies and can be used to purify intact inclusion bodies whose less soluble proteins can be extracted by other means.
B-PER complete bacterial protein extraction reagent
Thermo Scientific B-PER Complete Bacterial Protein Extraction Reagent is an all-in-one formulation that combines a lysis reagent with lysozyme and a universal nuclease to enable mild extraction from both gram-negative and gram-positive bacteria. Protein extracts are compatible with most protein assays and typical downstream applications such as affinity purification methods (e.g. GST, 6xHis) (Figure 5).
Figure 5. B-PER complete provides better compatibility with purification of 6His and GST fusion proteins. E. coli ER2566/pLATE51-Klenow, ER2566/pGST-CC-StpA and ER2566/pGST-Syk cell pellets cell pellets (0.5g), were resuspended in 2.5mL aliquots of B-PER Complete Reagent (B-PER) or competitor BugBuster master mix (Bug) with gentle vortexing for 15 minutes at room temperature. Insoluble cell debris was removed by centrifugation at 16,000 x g for 20 min at 4°C. 6xHis-Klenow protein was purified using Thermo Scientific HisPur Ni-NTA Agarose. GST-Syk and GST-StpA proteins were purified using Thermo Scientific Glutathione Agarose.
- C = control (E. coli lysate from before induction with IPTG)
- S = soluble protein (lysate after induction with 0.1mM IPTG)
- E = elution fraction after protein purification
Specialized lysis buffers are available for plants, yeast, and insects that contain specific enzymes to break down their cell wall and extract the soluble protein.
| Pierce Plant Total Protein Extraction Kit | Y-PER Yeast Protein Extraction Reagent | Y-PER Plus Dialyzable Yeast Protein Extraction Reagent | I-PER Insect Cell Protein Extraction Reagent | |
| When to use (compatible sample types) | Leaves, stems, soft roots, seeds | Saccharomyces cerevisiae, Schizosaccharomyces pombe and Pichia pastoris | Saccharomyces cerevisiae, Schizosaccharomyces pombe, Pichia pastoris, Bacillus subtilis, Escherichia coli | Suspended or adherent cultured insect cells (Sf9, Sf21) |
| Features | Obtain excellent protein yields in less than 10 minutes using a single buffer and a spin column | Does not require glass beads for extraction | Fully dialyzable formulation | Gentle, effective protein extraction without sonication |
| Compatible protein assays | BCA | BCA | BCA, Bradford assays | BCA |
| Downstream compatibility | SDS or native-PAGE, western blotting, immunoprecipitation, affinity purification, activity assays | SDS-PAGE, western blotting, protein purification, activity assays | SDS-PAGE, western blotting, protein purification | Western blotting, 6xHis-tagged protein purification, protein assays, ion-exchange chromatography |
| Available size(s) | 50 preps | 200 mL 500 mL | 500 mL | 250 mL |
| User guide | User Guide: Pierce Plant Total Protein Extraction Kit | User Guide: Y-PER Yeast Protein Extraction Reagent | User Guide: I-PER Insect Cell Protein Extraction Reagent | |
Plant tissue cell lysis kit
Plant cells are notoriously difficult to lyse and extract for proteomics work due to their tough cell walls and substantial polysaccharide content. The Thermo Scientific Pierce Plant Total Protein Extraction kit is composed of optimized buffers and filter cartridges that allow for efficient and rapid protein extraction in less than 10 minutes. The method has been validated for use with multiple plan organs (leaf, soft stem, root, seed, and flowers); multiple plant species (Arabidopsis, tobacco, maize, soybeans, peas, rice, and spinach); and fresh, frozen, and dehydrated tissue sources. The protein extracts can be used for applications such as SDS-PAGE, western blotting, immunoprecipitation, affinity purification, and activity assays.
Yeast cell lysis reagent
Thermo Scientific Y-PER Yeast Protein Extraction Reagent penetrates the tough yeast cell wall, perforating the cell wall and membrane and extracting soluble protein without completely damaging overall cell structure. Traditionally, yeast protein extraction required mechanical disruption with glass beads, making small-scale extraction difficult. However, Y-PER eliminated the need for glass bead lysis allowing for higher protein yield. Additionally, the elimination of glass bead methods allows for the protocol to be performed at room temperature and completed in 20 minutes.
Two formulations are available depending on the downstream application. Y-PER reagent is high salt (>300 mM) and is effective for S. cerevisiae, S. pombe, C. albicans, and P. pastoris (as well as various gram-positive and gram-negative bacteria). Although the detergent is compatible with various downstream methods, it is not dialyzable, so it cannot be removed in cases where incompatibility exists. Thermo Scientific Y-PER Plus Dialyzable Yeast Protein Extraction Reagent is a phosphate-free, low-salt formulation with a dialyzable detergent, validated for use primarily with S. cerevisiae.
Insect cell lysis reagent
Thermo Scientific I-PER Insect Cell Protein Extraction Reagent enables gentle extraction of soluble protein from baculovirus-infected insect cells grown in suspension of monolayer culture (both Sf9 and Sf21 cells). The reagent maintains functionality of extracted proteins and is directly compatible with downstream applications, such as protein assays, western blotting, and His-tagged protein purification (IMAC).
Nucleases are commonly added to cell lysis reagents to remove the viscosity caused by the DNA content in bacterial cell lysates or to remove the DNA templates from RNAs produced by in vitro transcription. Lysozyme increases protein or nucleic acid extraction efficiency by breaking down the bacterial cell wall.
| Pierce Universal Nuclease for Cell Lysis | DNase I Solution (2500 U/mL) | Micrococcal Nuclease Solution (≥100 U/µL) | Lysozyme | Lysozyme Solution | |
| Features | Degrades all forms of DNA and RNA | Degrades both single-stranded and double-stranded DNA | Degrades nucleic acids from cell lysates, releasing chromatin-bound proteins and shearing chromatin for use in chromatin immunoprecipitation (ChIP) experiments | Bacterial cell wall lytic enzyme that improves protein or nucleic acid extraction efficiency | |
| Available size(s) | 5 kU 100 kU 25 kU | 0.5 mL | 150 µL | 5 g | 50 mg/mL |
Cell lysis buffers for mammalian cells often contain detergents, salts, and other solutions to disrupt the lipid bilayer of the cell membrane and extract proteins, DNA, RNA, and other cellular components. These lysis buffers include the option to add protease or phosphatase inhibitors to aid in stabilizing the extracted sample.
Cell lysis buffers for mammalian culture cells
| RIPA Lysis Buffer | IP Lysis Buffer | M-PER Mammalian Protein Extraction Reagent | NP-40 Cell Lysis Buffer | Cell Lysis Buffer | |
| When to use | Extraction of cytoplasmic and nuclear protein in denaturing conditions | Formulated specially for pull-down and immunoprecipitation assays | Mild, non-denaturing and efficient lysis for cytoplasmic and nuclear protein extraction | Recommended for extraction of cytoplasmic proteins | Need a harsher buffer than NP-40 or when cytoplasmic and nuclear protein extraction is needed |
| Composition | 25 mM Tris-HCl, pH 7.6 150 mM NaCl 1% NP-40 1% sodium deoxycholate 0.1% SDS | A modified RIPA buffer formulation without SDS | Non-denaturing detergent in 25 mM bicine buffer (pH 7.6) | 50 mM Tris, pH 7.4 250 mM NaCl 5 mM EDTA 50 mM NaF 1 mM Na3VO4 1% NP-40 0.02% NaN3 | 10 mM Tris, pH 7.4 100 mM NaCl 1 mM EDTA 1 mM EGTA 1 mM NaF 20 mM Na4P2O7 2 mM Na3VO4 1% Triton X-100 10% glycerol 0.1% SDS 0.5% deoxycholate |
| Recommended supplemental protease/phosphatase inhibitor(s) | Halt Protease Inhibitor Cocktail | ||||
| Compatible protein assays | BCA assays, Detergent Compatible Bradford | BCA assays, Detergent Compatible Bradford | BCA assays, Bradford assays | BCA assays, Detergent Compatible Bradford | Detergent Compatible Bradford |
| Downstream compability | SDS-PAGE, western blotting | Immunoprecipitations, pull-down assays | IP, protein electrophoresis, western blots, ELISA, EMSA, reporter assays, purification, activity assays, amine reactive labeling | ELISA, protein electrophoresis, western blotting | ELISA, protein electrophoresis, western blotting |
| Available size(s) | 100 mL 250 mL | 100 mL 250 mL | 25 mL 250 mL | 100 mL | 100 mL |
| User guide | User Guide: RIPA Lysis and Extraction Buffer | User Guide: Pierce IP Lysis Buffer | User Guide: M-PER Mammalian Protein Extraction Reagent | User Guide: NP-40 Cell Lysis Buffer | User Guide: Cell Lysis Buffer |
Cell lysis buffers for mammalian tissues
| T-PER Tissue Protein Extraction Reagent | N-PER Neuronal Protein Extraction Reagent | |
| When to use (compatible sample types) | Heart, liver, kidney, lung, spleen | Brain tissue and primary neurons |
| Composition | Non-denaturing detergent in 25 mM bicine, pH 7.6 150 mM NaCl | -- |
| Compatible protein assays | BCA assays, Bradford assays | BCA assays, Detergent Compatible Bradford |
| Downstream compatibility | IP, western blots, ELISA, EMSA, purification, kinase assays, activity assays, amine reactive labeling | IP, pulldowns, western blots, ELISA, enzyme assays, amine reactive labeling |
| Available sizes | 500 mL | 100 mL |
| User guide | User Guide: T-PER Tissue Protein Extraction Reagent | User Guide: N-PER Neuronal Protein Extraction Reagent |
Mammalian cell lysis reagents for cultured cells
RIPA lysis buffer
RIPA (radioimmunoprecipitation assay) is a popular cell lysis option that allows for the extraction of membrane, cytoplasmic, and nuclear proteins from cells or tissues. This lysis buffer is particularly effective because of its non-ionic and ionic detergents such as SDS and sodium deoxycholate. Alternatively, these components may decrease activity of some protein kinases or enzymes, thus it is advised to prepare a RIPA buffer that does not contain SDS and sodium deoxycholate. Protease or phosphatase inhibitors can be utilized with RIPA buffer to prevent proteolysis and preserve the phosphorylation of proteins.
IP lysis buffer
Thermo Scientific Pierce IP lysis buffer is a whole cell lysis buffer based on a modified RIPA buffer formulation without SDS. This moderate-strength lysis buffer effectively solubilizes cellular proteins but does not liberate genomic DNA or disrupt protein complexes like ordinary RIPA buffer. IP lysis buffer is an alternative cell lysis option to RIPA lysis buffer when a protein affinity purification experiment, such as immunoprecipitation (IP) or co-immunoprecipitation (CoIP), is the downstream application (Figure 1). This lysis buffer was formulated specially for pull-down and immunoprecipitation assays when it is especially important that the lysis reagent does not contain denaturants or components that might interfere with the antibody binding or protein interactions of interest. Protease or phosphatase inhibitors can be used to prevent proteolysis and maintain the phosphorylation of proteins.
Figure 1. Immunoprecipitation of cell lysates extracted using different lysis buffers. A549 cell lysates (200µg) were generated using different lysis buffers. EGFR and PP2A were immunoprecipitated using the Thermo Scientific Pierce Crosslink IP Kit and anti-EGFR antibody (Stressgen) or anti-PP2A antibody (Millipore). The immunoprecipitations were washed with the same lysis buffer used to lyse the cells. Lysates representing 5% of the total load (L) and elution (E) fractions were blotted with anti-EFGR or anti-PP2A antibody and using Thermo Scientific SuperSignal West Dura Substrate for detection.
M-PER mammalian protein extraction reagent
Thermo Scientific M-PER Mammalian Protein Extraction Reagent is an effective yet milder alternative to RIPA buffer. This reagent can extract soluble protein from primary, suspension, and adherent cell types (Figure 2). Lysis can be completed in only 5 minutes and directly added in culture plates, requiring little to no additional cellular disruption. M-PER can result in significantly more extracted protein compared with freeze/thaw and sonication methods (data not shown). Cell lysates obtained using M-PER are compatible with several downstream assays including immunoassays, enzyme assays, and various reporter assays. Additionally, M-PER contains dialyzable components without primary amines, making it suitable for luciferase, beta-galactosidase, and CAT assays unlike alternative protein extraction methods.
Click image to enlarge
Figure 2. Protein yield from various cell types using M-PER Mammalian Protein Extraction Reagent. Cells were harvested at 85% confluency, washed twice and collected in ice-cold PBS, and counted. For each cell type, 1 x 106 cells were pelleted by centrifugation at 2,000 x g for 5 minutes and lysed in 1 mL M-PER Reagent for 5 minutes. The cell lysates were clarified by centrifugation at 14,000 x g for 10 minutes, the supernatant was collected and the protein concentration (μg/million cells) was determined using the Pierce BCA Protein Assay.
Mammalian cell lysis reagents for tissues
Thermo Scientific T-PER Tissue Protein Extraction Reagent is designed for total protein extraction from mammalian tissue samples, including heart, liver, kidney, lung, and spleen (Figure 3). The T-PER reagent utilizes a non-denaturing detergent in 25 mM bicine, 150 mM NaCl (pH 7.6), and is used in conjunction with mechanical or manual homogenization. Furthermore, T-PER reagent was validated for protein extraction from fresh and frozen tissues.
Thermo Scientific N-PER Neuronal Protein Extraction Reagent is designed for total protein extraction from mammalian neuronal tissue samples or primary cultured neurons. N-PER lysis can be completed in less than 30 minutes and is used in conjunction with mechanical or manual homogenization. Protease or phosphatase inhibitors can be added to prevent proteolysis or protect against phosphatase activity normally present in brain tissue.
Click image to enlarge
Figure 3. Protein extraction from various tissue types using T-PER Tissue Protein Extraction reagent. Duplicate tissue samples were weighed, resuspended in 1:10 to 1:20 w/v T-PER reagent, and disrupted in a chilled Dounce or benchtop tissue homogenizer. The resulting lysates were centrifuged at 10,000 x g for 5 minutes and the supernatant was collected. The protein concentration of each lysate was determined using the Pierce BCA Protein Assay to determine protein yield per milligram of starting tissue.
Click image to enlarge
Figure 4. Protein extraction from mouse brain neuronal tissue using N-PER Neuronal Protein Extraction reagent. Homogenization of mouse brain neuronal tissue in Thermo Scientific N-PER Reagent resulted in greater overall protein yield than in our standard T-PER Tissue Protein Extraction Reagent and two varieties of phosphate-buffered saline (PBS).
B-PER Bacterial Protein Extraction Reagents gently lyse E. coli and other species of bacterial cells and effectively extract soluble native and recombinant proteins. Several different ready-to-use formulations of B-PER reagent are available for different lysis needs. Depending on downstream application, protease inhibitors, salts, reducing agents, denaturants, and chelating agents may be added.
| B-PER Complete Bacterial Protein Extraction Reagent | B-PER Bacterial Protein Extraction Reagent | B-PER II (2X) Bacterial Protein Extraction Reagent | B-PER (PBS) Bacterial Protein Extraction Reagent | |
| When to use (compatible sample types) | E. coli, B. subtilis | E. coli, B. subtilis | E. coli, B. subtilis | E. coli, B. subtilis |
| Features |
|
|
|
|
| For use with fresh and frozen cells? | Yes, add EDTA to gram-negative fresh cells | Yes | Yes | Yes |
| Composition | B-PER reagent formulated in Tris buffer with enzymes (lysozyme & universal nuclease) | B-PER reagent formulated in Tris buffer | 2X B-PER reagent formulated in in Tris buffer | B-PER reagent formulated in PBS |
| Nuclease and lysozyme in formulation? | Yes | No | No | No |
| Sample processing time | 15 min | 10–15 min | 10–15 min | 10–15 min |
| Mechanical disruption required? | No | No, but for gram-positive bacteria add enzymes for increased yield | No, but for gram-positive bacteria add enzymes for increased yield | No, but for gram-positive bacteria add enzymes for increased yield |
| Amount of sample processed | 100 g cell paste per 500 mL | 125 g cell paste per 500 mL | 125 g cell paste per 250 mL | 125 g cell paste per 500 mL |
| Compatible protein assays | BCA, Bradford assays | BCA, Bradford assays | BCA, Bradford assays | BCA, Bradford assays |
| Downstream compatibility | SDS-PAGE, western blotting, protein purification | SDS-PAGE, western blotting, protein purification (except for GST fusion proteins) | SDS-PAGE, western blotting, protein purification (except for GST fusion proteins) | SDS-PAGE, western blotting, protein purification (except for GST fusion proteins) |
| Available size(s) | 250 mL 500 mL | 165 mL 250 mL 500 mL | 250 mL | 500 mL |
| User guide | User Guide: B-PER Complete Bacterial Protein Extraction Reagent | User Guide: B-PER Bacterial Protein Extraction Reagent | ||
B-PER Bacterial Protein Extraction Reagents
B-PER reagents can be used for various Gram-negative and Gram-positive bacteria such as S. aureus, H. pylori, and E. coli strains BL21(D3), JM109, DH5a, and M15. The reagent is also suitable for certain Archaebacteria species and cultured insect cells. Extraction does not require expensive equipment or mechanical disruption and can be performed in 10 to 15 minutes. B-PER reagent removes soluble protein from inclusion bodies and can be used to purify intact inclusion bodies whose less soluble proteins can be extracted by other means.
B-PER complete bacterial protein extraction reagent
Thermo Scientific B-PER Complete Bacterial Protein Extraction Reagent is an all-in-one formulation that combines a lysis reagent with lysozyme and a universal nuclease to enable mild extraction from both gram-negative and gram-positive bacteria. Protein extracts are compatible with most protein assays and typical downstream applications such as affinity purification methods (e.g. GST, 6xHis) (Figure 5).
Figure 5. B-PER complete provides better compatibility with purification of 6His and GST fusion proteins. E. coli ER2566/pLATE51-Klenow, ER2566/pGST-CC-StpA and ER2566/pGST-Syk cell pellets cell pellets (0.5g), were resuspended in 2.5mL aliquots of B-PER Complete Reagent (B-PER) or competitor BugBuster master mix (Bug) with gentle vortexing for 15 minutes at room temperature. Insoluble cell debris was removed by centrifugation at 16,000 x g for 20 min at 4°C. 6xHis-Klenow protein was purified using Thermo Scientific HisPur Ni-NTA Agarose. GST-Syk and GST-StpA proteins were purified using Thermo Scientific Glutathione Agarose.
- C = control (E. coli lysate from before induction with IPTG)
- S = soluble protein (lysate after induction with 0.1mM IPTG)
- E = elution fraction after protein purification
Specialized lysis buffers are available for plants, yeast, and insects that contain specific enzymes to break down their cell wall and extract the soluble protein.
| Pierce Plant Total Protein Extraction Kit | Y-PER Yeast Protein Extraction Reagent | Y-PER Plus Dialyzable Yeast Protein Extraction Reagent | I-PER Insect Cell Protein Extraction Reagent | |
| When to use (compatible sample types) | Leaves, stems, soft roots, seeds | Saccharomyces cerevisiae, Schizosaccharomyces pombe and Pichia pastoris | Saccharomyces cerevisiae, Schizosaccharomyces pombe, Pichia pastoris, Bacillus subtilis, Escherichia coli | Suspended or adherent cultured insect cells (Sf9, Sf21) |
| Features | Obtain excellent protein yields in less than 10 minutes using a single buffer and a spin column | Does not require glass beads for extraction | Fully dialyzable formulation | Gentle, effective protein extraction without sonication |
| Compatible protein assays | BCA | BCA | BCA, Bradford assays | BCA |
| Downstream compatibility | SDS or native-PAGE, western blotting, immunoprecipitation, affinity purification, activity assays | SDS-PAGE, western blotting, protein purification, activity assays | SDS-PAGE, western blotting, protein purification | Western blotting, 6xHis-tagged protein purification, protein assays, ion-exchange chromatography |
| Available size(s) | 50 preps | 200 mL 500 mL | 500 mL | 250 mL |
| User guide | User Guide: Pierce Plant Total Protein Extraction Kit | User Guide: Y-PER Yeast Protein Extraction Reagent | User Guide: I-PER Insect Cell Protein Extraction Reagent | |
Plant tissue cell lysis kit
Plant cells are notoriously difficult to lyse and extract for proteomics work due to their tough cell walls and substantial polysaccharide content. The Thermo Scientific Pierce Plant Total Protein Extraction kit is composed of optimized buffers and filter cartridges that allow for efficient and rapid protein extraction in less than 10 minutes. The method has been validated for use with multiple plan organs (leaf, soft stem, root, seed, and flowers); multiple plant species (Arabidopsis, tobacco, maize, soybeans, peas, rice, and spinach); and fresh, frozen, and dehydrated tissue sources. The protein extracts can be used for applications such as SDS-PAGE, western blotting, immunoprecipitation, affinity purification, and activity assays.
Yeast cell lysis reagent
Thermo Scientific Y-PER Yeast Protein Extraction Reagent penetrates the tough yeast cell wall, perforating the cell wall and membrane and extracting soluble protein without completely damaging overall cell structure. Traditionally, yeast protein extraction required mechanical disruption with glass beads, making small-scale extraction difficult. However, Y-PER eliminated the need for glass bead lysis allowing for higher protein yield. Additionally, the elimination of glass bead methods allows for the protocol to be performed at room temperature and completed in 20 minutes.
Two formulations are available depending on the downstream application. Y-PER reagent is high salt (>300 mM) and is effective for S. cerevisiae, S. pombe, C. albicans, and P. pastoris (as well as various gram-positive and gram-negative bacteria). Although the detergent is compatible with various downstream methods, it is not dialyzable, so it cannot be removed in cases where incompatibility exists. Thermo Scientific Y-PER Plus Dialyzable Yeast Protein Extraction Reagent is a phosphate-free, low-salt formulation with a dialyzable detergent, validated for use primarily with S. cerevisiae.
Insect cell lysis reagent
Thermo Scientific I-PER Insect Cell Protein Extraction Reagent enables gentle extraction of soluble protein from baculovirus-infected insect cells grown in suspension of monolayer culture (both Sf9 and Sf21 cells). The reagent maintains functionality of extracted proteins and is directly compatible with downstream applications, such as protein assays, western blotting, and His-tagged protein purification (IMAC).
Nucleases are commonly added to cell lysis reagents to remove the viscosity caused by the DNA content in bacterial cell lysates or to remove the DNA templates from RNAs produced by in vitro transcription. Lysozyme increases protein or nucleic acid extraction efficiency by breaking down the bacterial cell wall.
| Pierce Universal Nuclease for Cell Lysis | DNase I Solution (2500 U/mL) | Micrococcal Nuclease Solution (≥100 U/µL) | Lysozyme | Lysozyme Solution | |
| Features | Degrades all forms of DNA and RNA | Degrades both single-stranded and double-stranded DNA | Degrades nucleic acids from cell lysates, releasing chromatin-bound proteins and shearing chromatin for use in chromatin immunoprecipitation (ChIP) experiments | Bacterial cell wall lytic enzyme that improves protein or nucleic acid extraction efficiency | |
| Available size(s) | 5 kU 100 kU 25 kU | 0.5 mL | 150 µL | 5 g | 50 mg/mL |
Learn more: Traditional Methods of Cell Lysis
Ordering information
See all: Protein extraction buffers
Discover protein preparation tools
Protein sample preparation technical handbook
The Protein Preparation Handbook provides useful information on our broad selection of reagents and tools for protein extraction, clean-up, immunoprecipitation, and purification. Practical information, selection guides, and relevant data are included to help you improve your protein yield and downstream analysis.
Protein sample preparation eLearning course
This modular, animated, and narrated eLearning course was developed to provide a succinct, contextual summary of common laboratory methods and techniques required to achieve optimal results in protein assays and experiments. There are knowledge checks throughout the course to test what you have learned.
Related products and downstream applications
Stylesheet for Classic Wide Template adjustments
Stylesheet for Classic Wide Template adjustments
Hero banner dark gradient overlay
For Research Use Only. Not for use in diagnostic procedures.