GE Healthcare, PreScission Protease, 27-0843-01, One unit will cleave > 90% of 100 ?g of a test GST-fusion protein in 50 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 7.0 at 5°C for 16 h. Review

Let’s Dive into GE Healthcare, PreScission Protease, 27-0843-01, One unit will cleave > 90% of 100 ?g of a test GST-fusion protein in 50 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 7.0 at 5°C for 16 h.

As a seasoned protein purification specialist, I’ve seen my fair share of proteases promising efficient tag removal. But the GE Healthcare, PreScission Protease, 27-0843-01, One unit will cleave > 90% of 100 ?g of a test GST-fusion protein in 50 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 7.0 at 5°C for 16 h. boasts a unique combination of specificity and ease of use that truly piqued my interest. Manufactured by GE Healthcare, this protease offers a streamlined approach to GST-tag removal, crucial for downstream protein applications.

My quest for this particular protease stemmed from a recurring problem in our lab: inefficient and inconsistent tag cleavage. Previous enzymes often left behind residual tag fragments or required harsh conditions that compromised protein integrity. This PreScission Protease promised a cleaner, gentler, and more efficient solution for removing GST tags from our recombinant proteins.

The GE Healthcare PreScission Protease arrived in a well-protected vial, clearly labeled with concentration and storage conditions. My initial impression was positive; the liquid appeared clear and free of particulate matter, suggesting good purity.

Compared to trypsin or enterokinase, which I’ve used extensively, PreScission Protease offers site-specific cleavage, minimizing the risk of unintended proteolysis. I also considered AcTEV protease, but the ability to easily remove PreScission Protease itself post-cleavage via its GST-tag made it the more attractive option. Ultimately, the potential for efficient and controlled tag removal with minimal protein degradation swayed my decision.

I was cautiously optimistic about the PreScission Protease. Having been burned by underwhelming proteases in the past, I approached this one with a healthy dose of skepticism, hoping it would live up to its claims.

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Real-World Testing: Putting GE Healthcare, PreScission Protease, 27-0843-01, One unit will cleave > 90% of 100 ?g of a test GST-fusion protein in 50 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 7.0 at 5°C for 16 h. to the Test

First Use Experience

I first tested the PreScission Protease on a notoriously stubborn GST-fusion protein that consistently resisted complete cleavage with other enzymes. Following the recommended protocol, I incubated the protein with the protease in the specified buffer at 5°C for 16 hours. The low-temperature incubation helps maintain protein stability during the cleavage process.

The results were striking. After incubation, SDS-PAGE analysis showed virtually complete removal of the GST tag, with no signs of degradation or non-specific cleavage.

The enzyme’s activity remained consistent even with slight variations in buffer composition. This inherent robustness further streamlined downstream protein processing. There were no unexpected issues, only a sigh of relief at seeing such efficient cleavage.

Extended Use & Reliability

After several weeks of consistent use, the PreScission Protease has proven to be remarkably reliable. Cleavage efficiency remains high, and the enzyme shows no signs of losing activity.

Despite repeated freeze-thaw cycles (following recommended storage conditions), the enzyme has maintained its performance. Minimal handling is required to keep the enzyme in top condition. The PreScission Protease consistently outperforms previous enzymes in terms of both efficiency and consistency, streamlining our protein purification workflow and giving us confidence in our protein preparations.

Breaking Down the Features of GE Healthcare, PreScission Protease, 27-0843-01, One unit will cleave > 90% of 100 ?g of a test GST-fusion protein in 50 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 7.0 at 5°C for 16 h.

Specifications

The GE Healthcare, PreScission Protease, 27-0843-01, One unit will cleave > 90% of 100 ?g of a test GST-fusion protein in 50 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 7.0 at 5°C for 16 h. boasts the following specifications:

• Complete Packsize: 500 units. This provides a substantial quantity for numerous cleavage reactions, making it cost-effective for research labs.
• Application: Used for site-specific separation of the GST tag from proteins expressed using pGEX-6P vectors. Its specificity targets proteins created with specific vectors, streamlining processes for researchers.
• Unit Definition: One unit will cleave > 90% of 100 μg of a test GST-fusion protein in 50 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 7.0 at 5°C for 16 h. The standardized unit definition provides a clear benchmark for enzyme activity and ensures reproducibility.
• Vector: pGEX-6P-1, pGEX-6P-2, pGEX-6P-3. The protease’s compatibility with common pGEX vectors enhances its versatility and ease of integration into existing workflows.
• Molecular Weight: Mr 46000. Knowing the molecular weight is crucial for tracking the enzyme during purification and separation steps.
• Inhibitor: 100 mM ZnCl2 (< 50% inhibition); 100 μM chymostatin; 4 mM Pefabloc. Awareness of potential inhibitors helps avoid unintended inactivation of the protease.
• Elution Buffer: 50 mM Tris-HCl, 10 mM reduced glutathione, pH 8.0. Defining the elution buffer optimizes the protease removal step post-cleavage.
• Cleavage Buffer: 50 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM dithiothreitol (DTT), pH 7.0. Specifying the cleavage buffer ensures optimal protease activity and protein stability during tag removal.
• Supplied As: 2000 units/ml (833 to 1000 units/mg) in 50 mM Tris-HCl (pH 8.0), 150 mM NaCl, 10 mM EDTA, 1 mM DTT, and 20% glycerol. The supplied formulation ensures enzyme stability during storage and allows for easy dispensing and dilution.

These specifications are critical because they provide a clear understanding of the enzyme’s activity, specificity, and optimal operating conditions. Knowing these factors allows researchers to design experiments that maximize cleavage efficiency and minimize the risk of protein degradation.

Performance & Functionality

The GE Healthcare, PreScission Protease excels in its primary function: efficient and specific cleavage of GST tags. The enzyme consistently achieves >90% cleavage under the recommended conditions, resulting in clean separation of the target protein from the tag. Its high specificity minimizes off-target proteolysis, preserving the integrity of the protein of interest.

The protease’s ability to function at low temperatures is a significant advantage, allowing for cleavage reactions to be performed under conditions that promote protein stability. The primary drawback is the required 16-hour incubation time. While effective, it demands careful planning to align with project timelines. Overall, the enzyme consistently meets and often exceeds expectations, delivering reliable and high-quality results.

Design & Ergonomics

The protease is supplied in a user-friendly formulation that is easy to handle and dispense. The clear and concise product documentation further simplifies its use. No real learning curve exists; it can be readily incorporated into existing protein purification workflows.

Durability & Maintenance

The PreScission Protease is remarkably stable when stored according to the manufacturer’s recommendations. Repeated freeze-thaw cycles do not appear to significantly impact its activity. Minimal maintenance is required, simply aliquotting upon arrival. With proper handling, the enzyme can provide consistent performance for extended periods.

Accessories and Customization Options

The GE Healthcare, PreScission Protease doesn’t come with any specific accessories but integrates seamlessly with standard GST purification protocols. Post-cleavage, the protease itself can be easily removed using glutathione sepharose resin, thanks to its own GST tag. This capability eliminates the need for additional purification steps and ensures a clean protein product. It’s highly compatible with various downstream applications, like crystallography and structural biology studies.

Pros and Cons of GE Healthcare, PreScission Protease, 27-0843-01, One unit will cleave > 90% of 100 ?g of a test GST-fusion protein in 50 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 7.0 at 5°C for 16 h.

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Who Should Buy GE Healthcare, PreScission Protease, 27-0843-01, One unit will cleave > 90% of 100 ?g of a test GST-fusion protein in 50 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 7.0 at 5°C for 16 h.?

This protease is perfect for researchers and protein scientists who require efficient, specific, and reliable removal of GST tags from recombinant proteins. It’s particularly well-suited for those working with sensitive proteins that are prone to degradation or require native conformation.

If you’re on a tight budget or don’t mind compromising on cleavage efficiency, alternative and cheaper proteases may suffice. However, those aiming for the highest quality protein preparations will find the investment well worth it.

A must-have accessory for optimal use is glutathione sepharose resin, which allows for easy removal of the protease post-cleavage.

Conclusion on GE Healthcare, PreScission Protease, 27-0843-01, One unit will cleave > 90% of 100 ?g of a test GST-fusion protein in 50 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 7.0 at 5°C for 16 h.

The GE Healthcare, PreScission Protease is a powerful and reliable tool for GST-tag removal, offering a unique combination of efficiency, specificity, and ease of use. Despite the slightly higher price point and longer incubation time, the benefits in terms of protein quality and streamlined purification outweigh the drawbacks. I would wholeheartedly recommend it to any researcher seeking a top-tier protease for their protein purification needs.

If you’re tired of inconsistent cleavage and compromised protein integrity, give the GE Healthcare PreScission Protease a try. You won’t be disappointed.