Cell Lysis Buffers - US (2024)

Table of Contents

Mammalian cell lysis buffers for cultured cells Mammalian tissue cell lysis buffers Mammalian cell lysis reagents RIPA buffer M-PER mammalian protein extraction solution IP-compatible solutions Tissue protein extraction Bacterial cell lysis reagents B-PER solutions Insect, plant, and yeast cell lysis reagents Yeast cell lysis solutions Plant tissue cell lysis solutions Insect cell lysis solutions Nucleases and enzymes Product manuals Application notes and technical handbooks FAQs

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.

Click image to enlarge

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.

Click image to enlarge

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

User Guide: NP-40 Cell Lysis Buffer
User Guide: Cell Lysis Buffer
User Guide: RIPA Lysis and Extraction Buffer
User Guide: M-PER Mammalian Protein Extraction Reagent
User Guide: Pierce IP Lysis Buffer
User Guide: T-PER Tissue Protein Extraction Reagent
User Guide: N-PER Neuronal Protein Extraction Reagent

Bacterial

User Guide: B-PER Complete Bacterial Protein Extraction Reagent
User Guide: B-PER Bacterial Protein Extraction Reagent

Insect, plant and yeast

User Guide: I-PER Insect Cell Protein Extraction Reagent
User Guide: Pierce Plant Total Protein Extraction Kit
User Guide: Y-PER Yeast Protein Extraction Reagent

Application notes and technical handbooks

Protein extraction from neuronal tissue and primary cells
Protein sample preparation technical handbook
Western blot sample preparation protocol

FAQs

What are the buffers for cell lysis? ›

The most commonly used buffers are RIPA and NP-40 which are both suitable for whole-cell lysate/membrane bound proteins. RIPA buffer's harsh properties are best suited for hard to solubilize proteins, which is why it is the preferred choice for nuclear and mitochondrial proteins.

Learn More ›

How much lysis buffer to use for cells? ›

Add 200 to 500 µl of RIPA Lysis Buffer with Inhibitors to each plate and swirl to distribute buffer. If harvesting multiple plates of the same cell type, 0.5 to 1 ml of Lysis Buffer can be used to sequentially lyse at least 5 plates; this results in a higher concentration of protein in the final lysate.

View Details ›

What is the purpose of the lysis buffer solution? ›

A lysis buffer is a buffer solution used for the purpose of breaking open cells for use in molecular biology experiments that analyze the labile macromolecules of the cells (e.g. western blot for protein, or for DNA extraction).

Read On ›

What pH should my lysis buffer be? ›

7.5 to 7.6. The following can work well: lysis buffer containing 50 mM Tris•HCl, pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% Nonidet P-40, 5 mM Na3VO4, 20 mM NaF, 10 mM sodium pyrophosphate.

How to choose cell lysis buffer? ›

The main consideration when choosing a lysis buffer is whether the chosen antibody will recognize denatured samples. When this is not the case, it will be noted on the antibody datasheet, and buffers without detergent or with relatively mild non-ionic detergents (NP-40, Triton X-100) should be used.

Learn More Now ›

How do you lyse cells with lysis buffer? ›

To the cell pellet, add ice-cold PBS and wash the cells by centrifuging at 2,000 x g for 5-7 min at 4 °C. Add ice-cold lysis buffer to the cell pellet. Agitate the contents in microcentrifuge tubes for 30 min at 4 °C. Centrifuge the tubes at 16,000 x g for 20 min at 4 °C.

Read On ›

How long does lysis buffer take to work? ›

Glo Lysis Buffer (GLB), 1X, is a proprietary formulation developed to promote rapid lysis (within 5 minutes) of cultured mammalian cells without scraping or performing freeze-thaw cycles.

Discover More ›

How much ripa buffer for 1 million cells? ›

Add 200 µL of modified RIPA lysis buffer for every 1 million cells. 2. Boil at 95°C for 10 minutes as soon as possible after lysing. NOTE: It is important that the sample reaches 95°C.

How long can you keep lysis buffer? ›

If buffer will be continually used, it is recommended that the cell lysis buffer be kept at 4°C for 1-2 weeks. For longer periods of time, buffer should be stored at -20°C. Aliquotting of cell lysis buffer is recommended if many small experiments are to be performed.

Keep Reading ›

Does lysis buffer degrade DNA? ›

Firstly, it lyses the nuclear membrane in addition to a cell membrane. Secondly, it maintains the pH during the DNA extraction. Thirdly, it protects DNA from acidic degradation. Fourthly, it separates DNA from other cell debris.

What was the purpose of adding the lysis buffer to the cell culture? ›

Lysis buffer is then added to the resuspended cells in order to break them open and release cellular contents, including your plasmid of interest. (If interested, compare this protocol with a milder one used to extract proteins.) Lysis buffer contains SDS and NaOH, which serve to break bacterial cells open.

Keep Reading ›

How to store cell lysis buffer? ›

CST recommends that lysates are stored at -20℃ for no longer than 3 months. There are certain cell lines, treatments, and phosphorylation sites that are more sensitive to repeated freeze/thaw cycles. Make an effort to minimize your freeze/thaw cycles as much as possible.

How much cell lysis buffer to use? ›

Add 10 to 100 µl of RIPA Lysis Buffer with Inhibitors per 1 x 106 cells. The amount of lysis buffer should be empirically determined for each cell type to ensure efficient lysis as well as an optimal final concentration of protein in the lysate.

Learn More ›

At what pH do cells lyse? ›

At a pH range of 11.5–12.5, the OH⁻ ion reacts with the cell membrane, breaks the fatty acid-glycerol ester bonds, and subsequently makes the cell membrane permeable, and the SDS solubilizes the proteins and the membrane. The pH range of 11.5–12.5 is preferable for cell lysis [3,56].

What is cell lysis buffer? ›

The purpose of a cell lysis buffer is to use a chemical mixture to disrupt the exterior environment of a cell in a way that causes it to break open and release its contents.

Find Out More ›

What are the different types of lysis buffer for DNA extraction? ›

DNA can be extracted from cells using a variety of lysis buffers (and sometimes require mechanical methods). Reagents often used in lysis buffers include Tris, EDTA, SDS, CTAB, Triton X100, MgCl₂, KCl, NaCl and other detergents.

What is the formulation of cell lysis buffer? ›

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).

What are examples of buffers in cells? ›

Hemoglobin is one such example as it can bind hydrogen ions, especially prior to dissociation of oxygen. The other two main buffering systems in the blood and circulating cells are phosphates and the bicarbonate-carbonic acid buffer system. In the respiratory system, CO2 and carbonic acid are normally in equilibrium.

Get More Info ›

What is a cell lysis buffer for blood? ›

Pipette 1mL EDTA-treated whole blood into a tube containing 10-20 mL of Gibco™ ACK Lysing Buffer at room temperature. Allow the blood sample plus ACK Lysing Buffer to incubate at room temperature for 3 – 5 minutes. Lysis of the red cells should be evident during this incubation.