Effective cell lysis and protein extraction for different species of organisms and different cell and tissue types require different buffer formulations. Thermo Scientific and Invitrogen lysis buffers have been optimized and validated with specific tissue types, as well as in primary and cultured mammalian cells. Protein extracts are compatible with most protein assays and typical downstream applications. These convenient single formulation reagents typically allow samples to be processed in 5–10 minutes and offer high protein yields.
Mammalian cell lysis buffers for cultured cells
NP-40 Cell Lysis Buffer | Cell Lysis Buffer | M-PER Mammalian Protein Extraction Reagent | RIPA Lysis Buffer | IP Lysis Buffer |
When to use | ||||
Recommended for extraction of cytoplasmic proteins | Need a harsher buffer than NP-40 or when cytoplasmic and nuclear protein extraction is needed | Mild, non-denaturing and efficient lysis for cytoplasmic and nuclear protein extraction | Extraction of cytoplasmic and nuclear protein in denaturing conditions | Formulated specially for pull-down and immunoprecipitation assays |
Composition | ||||
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 | Non-denaturing detergent in 25 mM bicine buffer (pH 7.6) | 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 |
Compatible protein assays | ||||
BCA assays, Detergent Compatible Bradford | Detergent Compatible Bradford | BCA assays, Bradford assays | BCA assays, Detergent Compatible Bradford | BCA assays, Detergent Compatible Bradford |
Recommended applications | ||||
ELISA, protein electrophoresis, western blotting | ELISA, protein electrophoresis, western blotting | IP, protein electrophoresis, western blots, ELISA, EMSA, reporter assays, purification, activity assays, amine reactive labeling | SDS-PAGE, western blotting | Immunoprecipitations, pull-down assays |
Catalog No. | ||||
FNN0021 (100 mL) | FNN0011 (100 mL) | 78503 (25 mL) 78501 (250 mL) | 89900 (100 mL) 89901 (250 mL) | 87787 (100 mL) 87788 (250 mL) |
View recommended protein preparation buffers for mass spectrometry applications
Mammalian tissue cell lysis buffers
T-PER Tissue Protein Extraction Reagent | N-PER Neuronal Protein Extraction Reagent | |
---|---|---|
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 | |
Protein assay compatibility | 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 |
Catalog No. | 78510 (500 mL) | 87792 (100 mL) |
View recommended protein preparation buffers for mass spectrometry applications
Mammalian cell lysis reagents
RIPA buffer
RIPA buffer is a popular choice for lysis and protein extraction of mammalian cells or soft tissue. Originally named after the assay method for which it was developed (radioimmunoprecipitation assay), RIPA buffer is effective when the immediate downstream application is SDS-PAGE (denaturing polyacrylamide gel electrophoresis). The formulation includes two ionic detergents and one non-ionic detergent in Tris buffer: 25 mM Tris-HCl, pH 7.6, 150 mM NaCl, 1% NP40, 1% sodium deoxycholate, and 0.1% sodium dodecyl sulfate (SDS).
M-PER mammalian protein extraction solution
Thermo Scientific M-PER Mammalian Protein Extraction Reagent was developed as an effective yet milder alternative to RIPA buffer. M-PER reagent uses a non-denaturing detergent to prepare total cell lysate that is compatible with many downstream assays, including immunoassays, enzyme assays, and a variety of common reporter assays. Lysis can be performed directly in culture plates and is completed in only five minutes. Furthermore, significantly more protein can be obtained with this method compared with freeze/thaw and sonication.
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 (Cat. No. 23227).
IP-compatible solutions
When the immediate downstream application for a lysate is a protein affinity purification experiment, such as immunoprecipitation (IP) or co-immunoprecipitation (CoIP), it is especially important to ensure that the lysis reagent does not contain denaturants or components that might interfere with the antibody binding or protein interactions of interest. RIPA buffer, despite its name, is not always the best choice for immunoprecipitation experiments because it contains SDS. Our M-PER or IP Lysis Buffer are good alternatives. Thermo Scientific Pierce IP Lysis Buffer was formulated specially for pull-down and immunoprecipitation assays. IP Lysis Buffer is a mammalian whole cell lysis reagent 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.
Tissue protein extraction
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. 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. The resulting cell lysate, like the lysate prepared with M-PER reagent, is compatible with many functional assays.
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 (Cat. No. 23227) to determine protein yield per milligram of starting tissue.
Bacterial cell lysis reagents
Reagent | 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 |
Compatible sample types | E. coli, B. subtilis | E. coli, B. subtilis | E. coli, B. subtilis | E. coli, B. subtilis |
Composition | B-PER reagent formulated in Tris buffer with enzymes | 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 |
Works for fresh and frozen cells? | Yes, add EDTA to gram-negative fresh cells | Yes | Yes | Yes |
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 |
Protein assay compatibility | 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) |
Cat. No. | 89821 (250 mL) 89822 (500 mL) | 78243 (165 mL) 90084 (250 mL) 78248 (500 mL) | 78260 (250 mL) | 78266 (500 mL) |
B-PER solutions
Thermo Scientific B-PER Bacterial Protein Extraction Reagents gently lyse E. coli and other species of bacterial cells and effectively extract soluble native and recombinant proteins. B-PER reagents can be used for Gram-negative bacteria, 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 less than 10 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.
Several different ready-to-use formulations of B-PER reagent are available for different lysis needs. 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. These reagents provide high protein yield and typically allow samples to be processed in 10–15 minutes.
Insect, plant, and yeast cell lysis reagents
Reagent | Y-PER Yeast Protein Extraction Reagent | Y-PER Plus Dialyzable Yeast Protein Extraction Reagent | Pierce Plant Total Protein Extraction Kit | I-PER Insect Cell Protein Extraction Reagent |
Compatible sample types | Saccharomyces cerevisiae, Schizosaccharomyces pombe and Pichia pastoris | Saccharomyces cerevisiae, Schizosaccharomyces pombe, Pichia pastoris, Bacillus subtilis, Escherichia coli | Leaves, stems, soft roots, seeds | Suspended or adherent cultured insect cells (Sf9, Sf21) |
Features | Does not require glass beads for extraction | Fully dialyzable formulation | Obtain excellent protein yields in less than 10 minutes using a single buffer and a spin column | Gentle, effective protein extraction without sonication |
Protein assay compatibility | BCA | BCA, Bradford assays | BCA | BCA |
Downstream compatibility | SDS-PAGE, western blotting, protein purification, activity assays | SDS-PAGE, western blotting, protein purification | SDS or native-PAGE, western blotting, immunoprecipitation, affinity purification, activity assays | Western blotting, 6xHis-tagged protein purification, protein assays, ion-exchange chromatography |
Cat. No. | 78991 (200 mL) 78990 (500 mL) | 78999 (500 mL) | A44056 | 89802 (250 mL) |
Yeast cell lysis solutions
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. Y-PER reagent provides higher yields and greater flexibility for use in current proteomics workflows.
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.
Plant tissue cell lysis solutions
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.
Insect cell lysis solutions
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 and enzymes
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.
Reagent | Pierce Universal Nuclease for Cell Lysis | DNase I Solution (2500 U/mL) | Micrococcal Nuclease Solution (≥100 U/µL) | Lysozyme |
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 |
Cat. No. | 88700 (5 kU) 88702 (100 kU) 88701 (25 kU) | 90083 (0.5 mL) | 88216 (150 µL) | 90082 (50 mg/mL) 89833 (5 g) |
Product manuals
Mammalian
Bacterial
Insect, plant and yeast
Application notes and technical handbooks
Mammalian cell lysis buffers for cultured cells
NP-40 Cell Lysis Buffer | Cell Lysis Buffer | M-PER Mammalian Protein Extraction Reagent | RIPA Lysis Buffer | IP Lysis Buffer |
When to use | ||||
Recommended for extraction of cytoplasmic proteins | Need a harsher buffer than NP-40 or when cytoplasmic and nuclear protein extraction is needed | Mild, non-denaturing and efficient lysis for cytoplasmic and nuclear protein extraction | Extraction of cytoplasmic and nuclear protein in denaturing conditions | Formulated specially for pull-down and immunoprecipitation assays |
Composition | ||||
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 | Non-denaturing detergent in 25 mM bicine buffer (pH 7.6) | 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 |
Compatible protein assays | ||||
BCA assays, Detergent Compatible Bradford | Detergent Compatible Bradford | BCA assays, Bradford assays | BCA assays, Detergent Compatible Bradford | BCA assays, Detergent Compatible Bradford |
Recommended applications | ||||
ELISA, protein electrophoresis, western blotting | ELISA, protein electrophoresis, western blotting | IP, protein electrophoresis, western blots, ELISA, EMSA, reporter assays, purification, activity assays, amine reactive labeling | SDS-PAGE, western blotting | Immunoprecipitations, pull-down assays |
Catalog No. | ||||
FNN0021 (100 mL) | FNN0011 (100 mL) | 78503 (25 mL) 78501 (250 mL) | 89900 (100 mL) 89901 (250 mL) | 87787 (100 mL) 87788 (250 mL) |
View recommended protein preparation buffers for mass spectrometry applications
Mammalian tissue cell lysis buffers
T-PER Tissue Protein Extraction Reagent | N-PER Neuronal Protein Extraction Reagent | |
---|---|---|
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 | |
Protein assay compatibility | 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 |
Catalog No. | 78510 (500 mL) | 87792 (100 mL) |
View recommended protein preparation buffers for mass spectrometry applications
Mammalian cell lysis reagents
RIPA buffer
RIPA buffer is a popular choice for lysis and protein extraction of mammalian cells or soft tissue. Originally named after the assay method for which it was developed (radioimmunoprecipitation assay), RIPA buffer is effective when the immediate downstream application is SDS-PAGE (denaturing polyacrylamide gel electrophoresis). The formulation includes two ionic detergents and one non-ionic detergent in Tris buffer: 25 mM Tris-HCl, pH 7.6, 150 mM NaCl, 1% NP40, 1% sodium deoxycholate, and 0.1% sodium dodecyl sulfate (SDS).
M-PER mammalian protein extraction solution
Thermo Scientific M-PER Mammalian Protein Extraction Reagent was developed as an effective yet milder alternative to RIPA buffer. M-PER reagent uses a non-denaturing detergent to prepare total cell lysate that is compatible with many downstream assays, including immunoassays, enzyme assays, and a variety of common reporter assays. Lysis can be performed directly in culture plates and is completed in only five minutes. Furthermore, significantly more protein can be obtained with this method compared with freeze/thaw and sonication.
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 (Cat. No. 23227).
IP-compatible solutions
When the immediate downstream application for a lysate is a protein affinity purification experiment, such as immunoprecipitation (IP) or co-immunoprecipitation (CoIP), it is especially important to ensure that the lysis reagent does not contain denaturants or components that might interfere with the antibody binding or protein interactions of interest. RIPA buffer, despite its name, is not always the best choice for immunoprecipitation experiments because it contains SDS. Our M-PER or IP Lysis Buffer are good alternatives. Thermo Scientific Pierce IP Lysis Buffer was formulated specially for pull-down and immunoprecipitation assays. IP Lysis Buffer is a mammalian whole cell lysis reagent 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.
Tissue protein extraction
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. 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. The resulting cell lysate, like the lysate prepared with M-PER reagent, is compatible with many functional assays.
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 (Cat. No. 23227) to determine protein yield per milligram of starting tissue.
Bacterial cell lysis reagents
Reagent | 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 |
Compatible sample types | E. coli, B. subtilis | E. coli, B. subtilis | E. coli, B. subtilis | E. coli, B. subtilis |
Composition | B-PER reagent formulated in Tris buffer with enzymes | 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 |
Works for fresh and frozen cells? | Yes, add EDTA to gram-negative fresh cells | Yes | Yes | Yes |
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 |
Protein assay compatibility | 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) |
Cat. No. | 89821 (250 mL) 89822 (500 mL) | 78243 (165 mL) 90084 (250 mL) 78248 (500 mL) | 78260 (250 mL) | 78266 (500 mL) |
B-PER solutions
Thermo Scientific B-PER Bacterial Protein Extraction Reagents gently lyse E. coli and other species of bacterial cells and effectively extract soluble native and recombinant proteins. B-PER reagents can be used for Gram-negative bacteria, 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 less than 10 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.
Several different ready-to-use formulations of B-PER reagent are available for different lysis needs. 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. These reagents provide high protein yield and typically allow samples to be processed in 10–15 minutes.
Insect, plant, and yeast cell lysis reagents
Reagent | Y-PER Yeast Protein Extraction Reagent | Y-PER Plus Dialyzable Yeast Protein Extraction Reagent | Pierce Plant Total Protein Extraction Kit | I-PER Insect Cell Protein Extraction Reagent |
Compatible sample types | Saccharomyces cerevisiae, Schizosaccharomyces pombe and Pichia pastoris | Saccharomyces cerevisiae, Schizosaccharomyces pombe, Pichia pastoris, Bacillus subtilis, Escherichia coli | Leaves, stems, soft roots, seeds | Suspended or adherent cultured insect cells (Sf9, Sf21) |
Features | Does not require glass beads for extraction | Fully dialyzable formulation | Obtain excellent protein yields in less than 10 minutes using a single buffer and a spin column | Gentle, effective protein extraction without sonication |
Protein assay compatibility | BCA | BCA, Bradford assays | BCA | BCA |
Downstream compatibility | SDS-PAGE, western blotting, protein purification, activity assays | SDS-PAGE, western blotting, protein purification | SDS or native-PAGE, western blotting, immunoprecipitation, affinity purification, activity assays | Western blotting, 6xHis-tagged protein purification, protein assays, ion-exchange chromatography |
Cat. No. | 78991 (200 mL) 78990 (500 mL) | 78999 (500 mL) | A44056 | 89802 (250 mL) |
Yeast cell lysis solutions
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. Y-PER reagent provides higher yields and greater flexibility for use in current proteomics workflows.
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.
Plant tissue cell lysis solutions
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.
Insect cell lysis solutions
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 and enzymes
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.
Reagent | Pierce Universal Nuclease for Cell Lysis | DNase I Solution (2500 U/mL) | Micrococcal Nuclease Solution (≥100 U/µL) | Lysozyme |
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 |
Cat. No. | 88700 (5 kU) 88702 (100 kU) 88701 (25 kU) | 90083 (0.5 mL) | 88216 (150 µL) | 90082 (50 mg/mL) 89833 (5 g) |
Product manuals
Mammalian
Bacterial
Insect, plant and yeast
Application notes and technical handbooks
Related products and downstream applications
For Research Use Only. Not for use in diagnostic procedures.