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GE27-0843-01

PreScission Protease

Cytiva 27-0843-01

Synonym(s):

fusion protein, GST-tag removal

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About This Item

UNSPSC Code:
41106501
NACRES:
NA.56

packaging

pkg of 500 units

manufacturer/tradename

Cytiva 27-0843-01

shipped in

dry ice

storage temp.

−70°C

Related Categories

General description

PreScission Protease is a fusion protein of human rhinovirus (HRV) 3C protease and GST. It allows for on-column cleavage of GST tags and protein purification in one step.

On-column or off-column enzyme cleavage

GST tags and protease can be removed in a single step while fusion proteins are still bound to Glutathione Sepharose 4B, Glutathione Sepharose High Performance, or Glutathione Sepharose 4 Fast Flow affinity resin. Another option is to elute the GST-tagged protein using reduced glutathione, cleave, then pass the sample over the same resin to remove the protease and tags. Unlike other common proteases, such as thrombin and Factor Xa, this GST-tagged fusion protein can be removed easily through binding to a resin with affinity for GST. Other non-recombinant proteases require an additional purification step with a different resin.

Vectors encoding the HRV 3C protease recognition sequence

pGEX-6P vectors pGEX-6P-1, pGEX-6P-2, and pGEX-6P-3 encode the recognition sequence for this 3C protease.

Application

Specific, low temperature cleavage of fusion protein tags; Simple on-column cleavage during affinity purification
PreScission Protease has been used for eluting GST-tagged AT3, during in vitro ubiquitination reactions and AT3-Ub preparation. It has also been used to cleave GST tag from the a-enolase during induction of GST a-enolase expression in the protease-deficient BL21 strain of Escherichia coli.

Features and Benefits

  • Specific cleavage – between the Gln and Gly residues of the recognition sequence LeuGluValLeuPheGln/GlyPro.
  • Time-saving – GST-tagged proteins can be cleaved while still bound to an affinity resin.
  • Low incubation temperature – enables low temperature cleavage of fusion proteins containing the HRV 3C protease recognition sequence.

Storage and Stability

Please be aware this product may be shipped 90 days before the expiration date. For more information on the batch specific expiration date, please contact technical service.

Analysis Note

To view the Certificate of Analysis for this product, please visit www.cytiva.com.

Legal Information

pGEX Vectors
pGEX Vectors are to be used for scientific investigation and research and for no other purpose whatsoever and a license for commercial use of the licensed products and the processes claimed in US patent 5,654,176 and equivalent patents and patent applications in other countries must be negotiated directly with Millipore Corp (formerly Chemicon International Inc) by the purchaser prior to such use.

Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Zhen Wang et al.
Nature communications, 10(1), 3162-3162 (2019-07-20)
HECT E3 ligases control the degradation and functioning of numerous oncogenic/tumor-suppressive factors and signaling proteins, and their activities must be tightly regulated to prevent cancers and other diseases. Here we show that the Nedd4 family HECT E3 WWP1 adopts an
Dorothée Raoux-Barbot et al.
PloS one, 13(11), e0206133-e0206133 (2018-11-13)
Several bacterial pathogens produce nucleotidyl cyclase toxins to manipulate eukaryotic host cells. Inside host cells they are activated by endogenous cofactors to produce high levels of cyclic nucleotides (cNMPs). The ExoY toxin from Pseudomonas aeruginosa (PaExoY) and the ExoY-like module
Maximilian Fottner et al.
Nature communications, 12(1), 6515-6515 (2021-11-13)
The post-translational modification of proteins with ubiquitin (Ub) and Ub-like modifiers (Ubls) represents one of the most important regulators in eukaryotic biology. Polymeric Ub/Ubl chains of distinct topologies control the activity, stability, interaction and localization of almost all cellular proteins
Ana Belén Uceda et al.
Antioxidants (Basel, Switzerland), 10(2) (2021-03-07)
Frataxin is a mitochondrial protein which deficiency causes Friedreich's ataxia, a cardio-neurodegenerative disease. The lack of frataxin induces the dysregulation of mitochondrial iron homeostasis and oxidative stress, which finally causes the neuronal death. The mechanism through which frataxin regulates the
Sokol V Todi et al.
The Journal of biological chemistry, 285(50), 39303-39313 (2010-10-15)
Deubiquitinating enzymes (DUbs) play important roles in many ubiquitin-dependent pathways, yet how DUbs themselves are regulated is not well understood. Here, we provide insight into the mechanism by which ubiquitination directly enhances the activity of ataxin-3, a DUb implicated in
View All Related Papers

Articles

Troubleshooting in pGEX Expression Vectors

This page describes troubleshooting strategies for cloning the gene or gene fragment into a pGEX expression vector.

Troubleshooting of Cleavage Methods

This page shows troubleshooting strategies for cleavage methods using Cytiva products.

Cleavage and Purification of GST-Tagged Protein Bound to Glutathione Sepharose in Batch Mode

This page shows how to cleave and purify GST-tagged proteins bound to Glutathione Sepharose in batch mode from Cytiva. Glutathione Sepharose High Performance, Glutathione Sepharose 4 Fast Flow, and Glutathione Sepharose 4B can all be used for cleavage and purification of GST-tagged proteins in batch.

Cleavage and Purification of GST-Tagged Protein Bound to GSTrap

This page shows how to cleave and purify GST-tagged proteins bound to GSTrap from Cytiva.

See All

Protocols

Removal of Thrombin and Factor Xa Using HiTrap® Benzamidine FF (High Sub)

To protect the fusion protein from proteolytic degradation prior to enzymatic cleavage with PreScission Protease, thrombin, or Factor Xa, it may be necessary to remove proteases from the sample. Additionally, following enzymatic cleavage, it may be necessary to remove thrombin or Factor Xa from the sample.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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