Description
GoldBio's AEBSF (4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride) is a water-soluble, irreversible serine protease inhibitor used to prevent proteolytic degradation during protein extraction, purification, and sample preparation.
It rapidly inactivates a broad range of serine proteases, including trypsin, chymotrypsin, plasmin, kallikrein, thrombin, and plasma proteases, helping preserve protein integrity throughout analyses.
Unlike phenylmethylsulfonyl fluoride (PMSF), AEBSF is readily soluble in water and many aqueous buffers, generally avoiding the need for organic solvents during stock-solution preparation. Its improved stability in water and ease of use make it a convenient choice for cell lysis buffers, tissue homogenization, enzyme purification, immunoprecipitation, and other protein isolation work where minimizing proteolysis is essential.
AEBSF irreversibly inhibits serine proteases by covalently modifying the active-site serine residue, preventing enzymatic cleavage of target proteins.
Because AEBSF rapidly and irreversibly inactivates susceptible serine proteases, it is commonly added immediately before or during sample preparation to reduce protein degradation. Its compatibility with sensitive downstream analyses, particularly mass spectrometry, should be evaluated because AEBSF can also introduce covalent protein modifications.
- Serine protease inhibitor
- Inhibition of site-1 protease-dependent SREBP activation2
- Water-soluble alternative to PMSF/DFP
AEBSF has been used in cell culture in concentrations of up to 0.25mM. Aqueous solutions are stable between pH 5-6; limited stability above pH 7.5.
Attention Canadian and Mexican customers: This product is a hazardous and can only ship via Air.
Working Concentration
Make stock solutions at 100mM. Aliquot and store at -20°C for up to 2 months. AEBSF working concentrations are usually between 0.1-1.0mM and should be made daily.
Common Applications
(Click each for more information)
Protein Extraction and Cell Lysis
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Purpose: To prevent proteolytic degradation during protein extraction from cells and tissues.
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How It Works: AEBSF irreversibly inhibits serine proteases released during cell disruption, preserving protein integrity and improving recovery of intact proteins for downstream analysis.
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Applications: Cell lysis, tissue homogenization, protein extraction, Western blotting, and biochemical assays.
Wingfield, P. T. (2015). Overview of the purification of recombinant proteins. Current Protocols in Protein Science, 80, 6.1.1–6.1.35. https://doi.org/10.1002/0471140864.ps0601s80
Recombinant Protein Purification
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Purpose: To protect recombinant proteins from degradation during purification.
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How It Works: AEBSF may be added to compatible extraction and purification buffers to inhibit endogenous serine proteases that remain active during clarification and chromatography, helping reduce cleavage of susceptible recombinant proteins.
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Applications: Affinity chromatography, ion exchange chromatography, recombinant protein purification, and enzyme isolation.
Wingfield, P. T. (2015). Overview of the purification of recombinant proteins. Current Protocols in Protein Science, 80, 6.1.1–6.1.35. https://doi.org/10.1002/0471140864.ps0601s80
Preparation of Protease Inhibitor Cocktails
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Purpose: To provide broad protection against serine proteases as part of multi-component protease inhibitor formulations.
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How It Works: AEBSF can be combined with inhibitors targeting cysteine, aspartic, metalloprotease, or aminopeptidase activity to broaden protection against multiple protease classes during protein extraction and purification.
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Applications: Custom protease inhibitor cocktails, cell lysis buffers, tissue extraction, and proteomics sample preparation.
Ryan, C. A. (1990). Protease inhibitors in plants: Genes for improving defenses against insects and pathogens. Annual Review of Phytopathology, 28, 425–449. https://doi.org/10.1146/annurev.py.28.090190.002233
Proteomic Sample Preparation
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Purpose: To limit serine-protease activity during the early stages of protein extraction when the inhibitor can be removed or accounted for before mass-spectrometric analysis.
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How It Works: AEBSF can reduce post-lysis proteolysis during initial sample extraction, but excess inhibitor should be removed before trypsin digestion, and possible AEBSF-derived protein modifications should be considered during method development and data analysis.
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Applications: Early-stage proteomic sample extraction, protein-profile preservation before cleanup, and method development for samples requiring serine-protease inhibition.
Cai, C. X., Schneck, N. A., Zhao, W., Blackstock, D., Cai, J., Harris, D., ... & Lei, Q. P. (2019). Characterization of AEBSF-antibody modifications for a protease inhibitor supplementation strategy. Analytical and Bioanalytical Chemistry, 411(23), 6111–6118.
Key Benefits
· Water Soluble: Readily dissolves in water and many aqueous buffers, generally avoiding the organic-solvent stocks commonly required for PMSF.
· Irreversible Serine Protease Inhibition: Rapidly forms a covalent inhibitory complex with susceptible serine proteases, helping reduce protein degradation during extraction and purification.
· Broad Protease Coverage: Inhibits several well-characterized serine proteases, including trypsin, chymotrypsin, plasmin, kallikrein, and thrombin.
· Preserves Protein Integrity: Supports Protein Integrity: Helps reduce serine-protease-mediated degradation during cell lysis, protein purification, and sample preparation, supporting recovery of intact proteins.
· Compatible with Multiple Applications: Suitable for many cell lysis, protein purification, immunoprecipitation, and biochemical procedures when serine-protease inhibition is required. Compatibility with mass spectrometry and activity-based enzyme assays should be evaluated for each method.
· Convenient Alternative to PMSF: Provides broad serine-protease inhibition with greater aqueous solubility and improved solution stability relative to PMSF under compatible conditions.
Storage and Handling
Store at -20°C. Protect from light.
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