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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.

Simple Stop™ 1 is a broad-spectrum phosphatase inhibitor cocktail consisting of five phosphatase inhibitors in two solutions that target all the phosphatase categories: serine/threonine (Ser/Thr) specific phosphatase, tyrosine specific phosphatase, dual specificity phosphatases, acid phosphatase and alkaline phosphatase.

Each Simple Stop™ phosphatase inhibitor cocktail solution is a 100X concentrated, ready-to-use, solution. Simply add cocktail solution “A” and cocktail solution “B” to your extraction buffers or samples and you are ready to go. This cocktail is ideal for inhibition in tissue extractions and cell lysis experiments and is compatible with most common protein assays.

ProBlock™-100, EDTA Free is designed for large volumes of extraction/lysis buffers and is supplied as 10 vials per kit, with each vial suitable for 100 ml. ProBlock-100, EDTA Free protease inhibitor cocktail contains optimized concentrations of various protease inhibitors, which provide excellent inhibition of protease activities during protein purification. ProBlock™ is a superior general protease inhibitor cocktail that is suitable for purification from mammalian, plant, bacteria and yeast samples. The cocktail is EDTA free and contains both irreversible and reversible protease inhibitors to inhibit serine, cysteine and other proteases. At 1X concentration, ProBlock™ inhibits over 90% of protease activities (e.g. Mouse Pancreas Extract 0.5 mg/ml protein).

ProBlock™ Gold 2D is a protease inhibitor cocktail specifically developed for sample preparation for IEF/2D- studies. No EDTA is used, which allows optimal action of nuclease activity for removing nucleic acids from the samples.ProBlock™ Gold 2D contains both irreversible and reversible protease inhibitors and inhibits serine, cysteine and metalloproteases etc. Due to the optimized concentration of the various inhibitors, the ProBlock™ Gold 2D shows excellent inhibition of protease activities and is therefore suitable for the protection of proteins extracted from animal cells / tissues, plants, yeast and bacteria etc. ProBlock™ Gold 2D at 1X concentration in extraction buffer at pH 7-8 inhibits over 90% of protease activities.