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Proteinase K (one Column with 200 µl matrix G3m)

nr kat.: P3502
Opakowanie: 0.7mAnson U
Cena brutto: do wyceny
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Opis

Using our immobilized enzymes in compact reaction columns (CRC) saves your time in many molecular biological applications.
 
Features
  • Supplied in compact reaction columns (CRC)
  • 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
  • Reusability - 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
 
Description
  • 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 quick, 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.
 
Two different immobilization matrices are available:
 
  • Matrix F7m
    Matrix F7m has large pores. Molecules with up to 107 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.
  • Matrix G3m
    G3m has extremely small pores; this results in excellent recovery characteristics (i.e. complete recovery in very small volumes). Molecules larger than 103 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.
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Brief procedure in 4 steps:
  1. Equilibrate column with reaction buffer, since column is provided with storage buffer.
  2. Load substrate solution in reaction buffer.
  3. Incubate for 1 minute.
  4. Recover product solution by:
    a) centrifugation (smaller volumes)
    b) continuous flow mode (larger volumes)
 
Our 1 ml column »Mobicol«, specifically designed for this purpose has a lower plug and two screw caps. One cap has a Luer-lock adapter which can be closed. The enzymes are immobilized on 200 μl matrix which is placed between two filters.
The lower filters in the G3m columns have a pore size of 35 μm, in F7m columns the lower filters have 10 μm pore size. All upper filters have 90 μm pore size. The filter material is polyethylene.
The enzyme matrix is delivered in storage buffer. The volume above the upper filter is 800 μl. Reaction, washing and storage buffers are provided for the first applications. All buffers are specified on the detailed data sheets provided with the columns. These buffer conditions form the basis of our guarantee, however, many other combinations may work. Generally, immobilized enzymes also function under the conditions used for free enzyme reactions.
 
Specification
 
F7m:
  • 25 μg proteinase K per CR-column, immobilized on polyvinyl.
  • 0,7 mAnson units immobilized per CR-column.
  • This CR-column cuts at least 370 μg BSA per application.
  • Nr. 5 Storage buffer: 50 mM Tris/HCl, pH 7.5
  • Nr. 16 Reaction buffer: 50 mM Tris/HCl, 5 mM NaCl, pH 8.0
  • Nr. 17 Washing buffer: 50 mM Tris/HCl, 1.0 M NaCl, pH 8.0
  • The columns are more active in 0.1% SDS and at 40°C

 

MANUAL

 

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