TEV Protease

Product Name Cat. No. Size Conc.
TEV Protease PR002S 1,000 unit 10 unit/μl
TEV Protease PR002M 2,000 unit 10 unit/μl
TEV Protease PR002L 5,000 unit 10 unit/μl
리뷰 0 위시 0

상품간략정보 및 구매기능

TEV Protease

상품 선택옵션 0 개, 추가옵션 0 개

제조사 PR002L
원산지 5,000 unit
브랜드 10 unit/μl
판매가격 1,200,000원
배송비결제 주문시 결제
  • TEV Protease
    +0원

Product description

This product is a recombinant sequence-specific protease purified to near homogeneity from E. coli that expresses a gene encoding TEV Protease of Tobacco Etch Virus. TEV Protease recognizes a 7 amino acid sequence (EXXYXQ ↓ G/S) and cleaves the peptide bond between glutamine and glycine or serine (indicated by an arrow ↓).

Characteristics
- Molecular weight : 28.4 kDa
- Reaction temperature : 4 ~ 30℃
- Optimal pH range : pH 5.5 ~ 8.5 (optimal pH=7)

Applications
- Removal of fusion tag from recombinant proteins

Quality control
- Purity : >95% on SDS-PAGE
- Non-specific protease-free
- Endonuclease-free
- Exonuclease-free
- RNase-free

Unit definition
One unit of TEV Protease is the amount of enzyme required to cleave >85% of 3 μg of substrate protein in 1 hr at 30℃ in the 1X TEV Protease buffer.

Storage buffer
50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 5 mM DTT, 0.1% TritonX-100, 50% glycerol

10X TEV Protease buffer

500 mM Tris-HCl (pH 8.0), 5 mM EDTA, 1 mM DTT 

 

Cautions

- It is important to confirm cleavage efficiency of TEV Protease with each substrate, since it varies greatly depending on the type of substrates used, incubation temperature, incubation period, and the amount of TEV Protease used. Some substrates, despite the presence of a cleavage site, may not be cleaved if the site is not accessible to the enzyme. This can be overcome by inserting polyglycine, polyhistidine, or a FLAG-tag epitope next to a TEV Protease cleavage site. This allows the recognition sequence to be exposed to the TEV Protease.

- After cleavage reaction, TEV Protease can be efficiently removed with a Ni2+-affinity column due to the presence of a polyhistidine tag at its N-terminus. Prior to a Ni2+-affinity column, EDTA and DDT should be removed. The addition of 10 mM Mg2+ can effectively neutralize EDTA. Generally, substrate proteins bound on the affinity resins in a column requires a larger amounts of TEV Protease than proteins in solution. 

 

Figure 1. Specific cleavage of substrates by TEV Protease.

Three different substrate proteins (3 μg each) that include MBP-GST, bovine serum albumin (BSA), and mutated MBP-GST (TEV-cleavage site mutant) were incubated at 30℃ for 1 hr with 10 units (1.8 μg) of TEV protease (Cat.# PR002). Only the MBP-GST substrate that contains a correct TEV cleavage site (ENLYFQ^G) present between GST (glutathione S-transferase) and MBP (maltose binding protein) was cleaved into MBP and GST. Neither BSA nor the mutated MBPGST was not cleaved by TEV protease. The mutated MBP-GST contained an amino acid change in the cleavage site. + and - indicate the additions and omissions, respectively, of TEV protease in reaction mixtures. Migration positions of proteins used or products formed were indicated by arrows. Mw, molecular weight marker.

Certificates of analysis

  • 등록된 내용이 없습니다.

Products related to the selected product.

선택하신 상품이 장바구니에 담겼습니다.

계속구매 장바구니 이동
  • Add.

    281-9, Munji-ro, Yuseong-gu, Daejeon, 34050, Republic of Korea

  • Tel. +82 - 42 - 719 - 1023
  • Fax. +82 - 42 - 719 - 1024
  • Email. info@enzynomics.com

copyright© Enzynomics co Ltd, All rights reserved.

상단으로