Description
GoldBio's ProBlock™ Gold Extra Strength Protease Inhibitor Cocktail is a concentrated, broad-spectrum inhibitor formulation designed for demanding protein extraction and purification applications where increased protease protection is needed.
It contains 50% higher levels of protease inhibitors than the standard ProBlock™ Gold formulation and combines reversible and irreversible inhibitors targeting serine, cysteine, aspartic, aminopeptidase, and other protease activity.
At a 1X concentration in extraction buffer at pH 7 to 8, ProBlock™ Gold Extra Strength inhibits more than 90% of protease activity in GoldBio's mouse pancreas extract test system.
The disclosed formulation contains AEBSF, aprotinin, bestatin, E-64, leupeptin, pepstatin A, and PMSF. Combining inhibitors with different target specificities provides broader protection than relying on a single protease inhibitor.
The Extra Strength formulation is especially useful for samples with high endogenous protease activity or when maximizing recovery of intact proteins during initial extraction and purification is important.
ProBlock™ Gold Extra Strength is supplied with separate EDTA and 1,10-phenanthroline monohydrate solutions for optional metalloprotease inhibition. Researchers can add either chelator when additional metalloprotease protection is needed or leave them out when metal chelation could interfere with the experiment.
This is particularly important for proteins that require Ca²⁺, Mg²⁺, or Mn²⁺ for biological activity and for immobilized metal affinity chromatography. Omitting EDTA and 1,10-phenanthroline can also improve compatibility with applications such as isoelectric focusing.
Components
- ProBlock™ Gold Extra Strength Protease Inhibitor Cocktail
- EDTA (0.5 M Solution, pH 8.0)
- 1,10-Phenanthroline monohydrate (1 M Solution [100X])
Cocktail Protease Inhibitors:
- AEBSF
- Aprotinin
- Bestatin
- E-64
- Leupeptin
- Pepstatin A
- PMSF
Mechanism
ProBlock™ Gold Extra Strength uses a multi-inhibitor approach to provide increased protection against proteolysis in samples with elevated or difficult-to-control protease activity. Its higher inhibitor content provides increased inhibitor availability during protein extraction and purification from samples with substantial protease activity. The combination targets multiple protease classes simultaneously, reducing the likelihood that active endogenous proteases will continue degrading proteins after cells or tissues are disrupted.
Comparison
|
Feature
|
ProBlock™ Gold Extra Strength
(Catalog Number: GB-116)
|
ProBlock™ Gold [100X]
(Catalog Number: GB-108)
|
|
Best For
|
More demanding protein extraction and purification where increased inhibitor strength is desired
|
Routine protein extraction and purification
|
|
Core Inhibitors
|
AEBSF, aprotinin, bestatin, E-64, leupeptin, pepstatin A, and PMSF
|
AEBSF, aprotinin, bestatin, E-64, leupeptin, pepstatin A, and PMSF
|
|
Protease Coverage
|
Broad-spectrum inhibition with an enhanced dose of protease inhibitors
|
Broad-spectrum inhibition of serine, cysteine, and other proteases
|
|
Metalloprotease Inhibition
|
Separate EDTA and 1,10-phenanthroline monohydrate solutions included
|
Separate EDTA solution included
|
|
EDTA
|
Optional
|
Optional
|
|
1,10-Phenanthroline
|
Included separately for additional metalloprotease inhibition
|
Not included
|
|
Metal-Sensitive Applications
|
EDTA and 1,10-phenanthroline can be omitted when divalent cations must be preserved
|
EDTA can be omitted when divalent cations must be preserved
|
|
Storage
|
4°C
|
4°C
|
|
Stability
|
At least 1 year when stored and handled as recommended
|
At least 1 year when stored and handled as recommended
|
Common Applications
(Click each for more information)
Protein Purification from Protease-Rich Samples
-
Purpose: To provide increased protection against protein degradation when routine protease inhibition may not provide sufficient coverage.
-
How It Works: The Extra Strength formulation contains 50% more protease inhibitors than standard ProBlock™ Gold. Its combination of AEBSF, aprotinin, bestatin, E-64, leupeptin, pepstatin A, and PMSF targets multiple protease classes simultaneously.
-
Applications: Protein purification from protease-rich samples, difficult tissue extracts, purification of proteolytically sensitive proteins, and experiments where maximizing recovery of intact protein is important.
Preparation of Lysates for Quantitative Protein Assays
-
Purpose: To preserve proteins during tissue extraction before quantitative analysis of specific protein targets.
-
How It Works: Adding the inhibitor cocktail during homogenization limits proteolytic changes that could alter the amount or integrity of proteins subsequently measured. This is particularly useful when samples will be stored before analysis.
-
Applications: ELISA, cytokine quantification, comparative protein analysis, and preparation of frozen lysates for later measurement.
Romero, J., Toral-Rios, D., Yu, J., Paul, S. M., & Cashikar, A. G. (2024). 25-hydroxycholesterol promotes brain cytokine production and leukocyte infiltration in a mouse model of lipopolysaccharide-induced neuroinflammation. Journal of Neuroinflammation, 21, 251. https://doi.org/10.1186/s12974-024-03233-1
Benefits:
- Greater Protease Inhibition: Provides >90% protease inhibition at 1X concentration and pH 7–8 in GoldBio's mouse pancreas extract test system.
- Contains 50% higher levels of protease inhibitors than standard ProBlock™ Gold.
- Supports Recovery of Intact Proteins: Increased inhibitor levels help reduce proteolytic degradation during initial extraction and purification, supporting recovery of intact target proteins from protease-rich samples.
- Disclosed Formulation: Contains AEBSF, aprotinin, bestatin, E-64, leupeptin, pepstatin A, and PMSF.
- Multiple package sizes
Storage/Handling:
Store at 4°C. When stored under the recommended conditions and handled correctly, these products should be stable for at least 1 year from the date of receipt.
Powered by Bioz