Update profile
    Change password
    Sign out

TPCK Trypsin and Immobilized TPCK Trypsin


Cleaves at the carboxy-side of arginine and lysine.

Pierce TPCK Trypsin

Thermo Scientific Pierce TPCK Trypsin is a serine endoprotease that is applicable to amino acid analysis and protein sequencing, mapping and structural studies; the immobilized form allows sample separation after treatment.

Trypsin has a wide range of applications including amino acid analysis and protein sequencing, mapping and structural studies. Enzymes such as trypsin and chymotrypsin have become important tools in sequencing studies since they are highly selective in their cleavage of peptide bonds. Trypsin cleaves only those peptide bonds in which the carboxyl group is contributed by a lysine or an arginine residue, regardless of the length or amino acid sequence of the chain. Our Immobilized TPCK Trypsin can be substituted for free trypsin in many application and is advantageous because it minimizes autolysis, eliminates contamination of a sample with the protease and allows control of the digestion by removing the trypsin. Immobilized trypsin is also more stable against heat-induced denaturation, resulting in longer maintenance of activity.

Applications:

  • Removal of adherent cells from tissue culture flasks
  • Preparing tryptic fragments for Edman degradation sequencing
  • Immobilized trypsin can be used to purify soybean trypsin inhibitor
  • Use our Pierce Trypsin, MS Grade (Part No. 90057) for mass spectrometry workflows

Product Details:

Trypsin is a 23.8kDa pancreatic serine endoprotease derived from trypsinogen, an inactive precursor zymogen, after enzymatic removal of an n-terminal leader sequence by enterokinase. Once some trypsin has been formed it can catalyze the conversion of more trypsinogen into its catalytically-active form. Trypsin is treated with L-1-tosylamido-2-phenylethyl chloromethyl ketone (TPCK) to inhibit contaminating chymotrypsin activity without affecting trypsin activity.

Properties of Trypsin Protease.
Specificity Cleaves specifically at the carboxyl side of arginine and lysine residues
Source Bovine pancreas
Molecular weight 23,800
Protease type Serine protease
Uses/applications Specific protein fragmentation for protein identification and sequence analysis
Reaction conditions pH 7.5-9.0, 37°C
Storage conditions Store at 4°C
Inhibitors TLCK, DFP, Aprotinin, PMSF
Unit definition One unit is equal to 1 μmole of TAME (p -toluenesulfonyl-L-arginine methyl ester) in the presence of Ca2+ hydrolyzed/minute at pH 8.2, 25°C. (One TAME unit = 57.5 National Formulatory Units).

References:

  1. Cunningham, L.W. (1954) Molecular-kinetic properties of crystalline diisopropyl phosphoryl trypsin. J. Biol. Chem. 211, 13.
  2. Walsh, K.A. and Neurath, H. (1964) Trypsinogen and chymotrypsinogen as homologous proteins. Proc.Natl. Acad Sci. USA, 52, 884.
  3. Kostka, V. and Carpenter, F.H. (1964) Inhibition of chymotrypsin activity in crystalline trypsin preparations. J. Biol. Chem. 239, 1799-1803.
  4. Walsh, K.A. "Trypsinogens and trypsins of various species" in Methods of Enzymology, Vol. XIX, (Perlman, G. E; and Lorand, L; eds) pp. 41.
  5. Woolhead , C.A., et al. (2001) J. Biol. Chem. 276, 14607-14613.
  6. Rush, J., et al. (2005) Nature Biotech. 23, 94-101.

Related Products:

Pierce Trypsin, MS Grade (Part No. 90057)
MS-Grade Proteases and Digestion Kits for Mass Spectrometry
Standard-Grade Proteases and Protease Resins
Enzymes for Cell Lysis


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
20230 Immobilized Trypsin, TPCK Treated
Formulation: Trypsin on crosslinked 4% beaded agarose, slurried in 50% glycerol containing sodium azide
Sufficient For: At least 200 TAME units of activity/mL resin
2mL Product Instructions for product #20230 Immobilized Trypsin, TPCK Treated MSDS for product #20230 Immobilized Trypsin, TPCK Treated Certificate of Analysis for product #20230 Immobilized Trypsin, TPCK Treated $78.00

Add to cart
20233 TPCK Trypsin
Formulation: Lyophilized enzyme powder
Sufficient For: At least 180 TAME units/mg
50mg MSDS for product #20233 TPCK Trypsin Certificate of Analysis for product #20233 TPCK Trypsin $73.00

Add to cart

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

Share This Page
 
 

Follow Us
  Email Sign-up  Email Announcements

 



Antibodies  |   Molecular Biology   |   Cell Biology   |  Thermo Scientific  |   * Trademarks  |   Privacy Statement
© 2014 Thermo Fisher Scientific Inc.   |   3747 N Meridian Rd, Rockford, IL USA 61101   |   1-800-874-3723  or  815-968-0747