Protease Pro39

Cat No	Y1-100-005	Y1-100-025
Qty	500 U	2500 U
Source	Recombinant E.coli strain
Storage	- 20 oC
Storage buffer	Contains 40% glycerol

  Enzyme properties

• Specific cystein protease
• Used for cleavage of fusion proteins 
• Cleaves both in liquid as well as in a resin-bound state
• Cleaves amino acid sequence DVLRLGRAGA/G  at the N-terminus or DVLRLGRAGA/XIFSS at the C-terminus, where X is preferably Y or G but, can be any amino acid except P
  

Product description Pro39 is the Semliki Forest Virus (SFV) non-structural protein nsP2 C-terminal protease domain, in a truncated, modified and His-tagged form. Pro39 protease specifically recognizes and cleaves the amino acid sequence DVLRLGRAGA/YIFSS or DVLRLGRAGA/G . The cleavage is highly specific and active. The molecular weight of Pro39 is approximately 39 kDa. Since the protease has a His-tag, it can be easily removed from cleavage reactions. The reaction conditions for cleaving of fusion proteins have to be determined empirically. During cleavage reactions, it is recommended that samples be removed from the reaction mixture at various points in time and analyzed by SDS-PAGE to estimate the yield and extent of digestion, and the purity of the product. Typical parameters that should be varied in pilot experiments include enzyme/substrate ratio, substrate concentration, temperature, reaction time, pH. Characteristics

• Pro39 cleaves substrates in a wide temperature range +4...+ 36ºC (optimal is +30ºC), in neutral pH conditions (i.e. pH 7-8) in most commonly used buffer systems, including Tris-HCl, HEPES and phosphate buffers.
• Pro39 cleaves substrates in the presence of high concentration NaCl (up to 4M) or urea (up to 1M).
• The cleavage of the substrates can be reversibly inhibited by the addition of Zn-ions and reactivated by the addition of EDTA
• Pro39 cleaves substrates having the recognition site both in liquid
• Inhibitors:
  • divalent cations (Ni, Co, Cd, Zn)
  • 0,1% SDS
  • 0,5M guanidine
  • N-ethylmaleimide
• Pro39 is resistant to the inhibitors of serine proteases (PMSF), metalloproteases (EDTA), aspartic proteases (pepstatin), and some cystein protease inhibitors (leupeptin and E-64)

Unit definition
1 unit Pro39 will cleave 10 µg of test substrate to 90% completion in 1 hour at 30ºC Optimal reaction conditions
Buffer: 50mM Tris-HCl (pH 7.5), 150mM NaCl, 1mM EDTA, 1mM DTT
Temperature: + 30ºC