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Immobilized Enzymes

P3302 - alpha-Chymotrypsin immobilized on matrix G3m, 1 column

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DESCRIPTION: alpha-Chymotrypsin immobilized on 200ul of matrix G3m, one Compact Reaction Column (CRC). Contains 1.4 units of alpha-chymotrypsin. The enzyme is TLCK treated and therefore free of trypsin activity. It hydrolyzes peptides, amides, and esters preferentially at the carboxyl groups of hydrophobic amino acids (L-tyrosine, L-phenylalanine, and L-tryptophan but also bonds of leucyl, methionyl, asparaginyl, and glutamyl residues).

ADVANTAGES OF IMMOBILIZED ENZYMES FOR SOLID PHASE ENZYME REACTIONS
Using our immobilized enzymes in compact reaction columns (CRC) saves your time in many molecular biological applications.

Features:

  • high enzyme concentration and activity
  • skip extensive steps in lab protocols
  • convenience and ease of handling
  • omit phenol extractions
  • avoid contaminating the lab (and your reaction solutions) with enzymes (RNase!)
  • enjoy short reaction and treatment times
  • get absolutely clean solutions
  • re-usability - use CR columns over and over again
  • take CR columns even for tiny solution volumes (20 µl!)
  • retain small volumes even after treatment
  • treat larger volumes using syringes or tubing with standard Luer-lock connections

GENERAL PRODUCT DESCRIPTION:p>Compact reaction columns are small volume columns (Mobicols) containing a matrix to which enzymes are bound (immobilized covalently). Enzyme reactions occur when the substrate is added to the column. The sample is recovered from the column quantitatively by washing or centrifugation. The enzyme remains bound to the column.

Our Immobilization Technology was developed in co-operation with the Max Planck Society and is patented. It enables us to offer enzymes immobilized on both polyvinyl and dextran matrix. The immobilized enzymes retain their full activity and possess an extremely high occupational density.

Immobilized enzymes are supplied in extremely versatile compact reaction columns (CRC) which fit into 1.5 ml microcentrifuge tubes. The columns have Luer-lock fittings, allowing direct syringe application of substrate solution, continuous flow processing of bulk solutions, or application of pressure for recovering the substrate. For small substrate volumes (approx. 50 µl or less), most enzyme columns can be spun dry in benchtop centrifuges for fast, effective recovery.

The immobilized enzymes are extremely stable in aqueous media over a pH range of 5 to 10 and column bleeding is negligible. The "stiff" linkers which separate the enzyme from the matrix surface effectively eliminate steric hindrance. As a result, the enzymes retain essentially their full activity in the immobilized state.

This well established technology allows you to expose your reaction solutions to very high concentrations of modifying enzymes. The exposure to high enzyme concentrations allows reaction times to be greatly reduced.

CHOICE OF IMMOBILIZATION MATRIX:

Two different immobilization matrices are available: F7m and G3m. They have different pore characteristics:

* Matrix F7m has large pores. Molecules with up to 10↑7 Dalton molecular weight (most enzymes and substrates) can enter these pores. The total surface of the material is very large, resulting in an extremely high enzyme activity on the matrix.

* G3m has extremely small pores; this results in excellent recovery characteristics (i.e. complete recovery in very small volumes). Molecules larger than 10↑3 Dalton molecular weight (larger peptides, proteins and nucleic acids) cannot enter these pores. The total surface area of the material is smaller than for F7m resulting in a smaller enzyme activity on the matrix.

Your first choice should be F7m, since it is highly active. The product is washed out of the column with an additional 200ul-500ul of buffer. However if you work with small volumes and cannot tolerate dilution of your sample solution by 200ul, then you should use G3m. The G3m columns are less active, but you can spin all of your product solution out of the column with high yield and negligible dilution