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Collagenase & Neutral Protease
Collagenases – For Tissue Dissociation
Collagenases from Clostridium histolyticum are proteolytic enzymes that
are able to cleave peptide bonds in the triple helical collagen molecule of
human or animal tissue in situ.
The collagenases are divided into
class I and class II collagenase isoforms on the basis of their activities
towards synthetic peptides. Both collagenase isoforms act on triple helical
type I, II, III and IV collagens, but in slightly different modes of action.
SERVA Collagenase NB qualities are particularly suitable for various
tissue isolation applications. The application of pharmaceutical manufacturing
standards guarantees stringent quality control, high quality, reliable
lot-to-lot consistency and excellent performance of the product.
TSE safety
of the fermentation product has been certified by the European Authorities,
EDQM and by supplier’s certificate of origin.
Reference Collagenases N (cat. no. 17463, 17464) are available for
enzyme activity determination or incoming goods inspection according to cGMP.
Anti-Collagenase polyclonal antibody from sheep (cat. no. 58050) can be
used in ELISA and Western blot experiments, immunostaining of tissue and
analysis of residual collagenase impurities in cell preparations.
Stability and Storage
Collagenase requires calcium ions both for full catalytic activity and binding
to the collagen molecule. Collagenase NB qualities are soluble in water and
diluted salt solutions. The enzyme is reversibly inactivated at high pH values
and irreversibly inactivated at low pH values.
Inhibitors of collagenase
include cysteine, EDTA, o-phenanthroline, 8-hydroxyquinoline-5-sulfonate,
bipyridyl and 2,3-dimercaptopropanol. It is also inhibited by TRIS buffer
above pH 7.5. Collagenase is not inhibited by diisopropylphosphorofluoridate.
All
Collagenase NB products delivered as lyophilized powders can be stored at +2
to +8 °C and remain stable without loss of activity for at least three years.
Enzymes should be protected from moisture.
Dissolved enzymes can be
aliquoted and stored at -20°C for one year. Repeated freezing and thawing
should be avoided.
Conversion rates of collagenase activity units
SERVA
states collagenase activity in PZ units according to Wünsch:
1 PZ U/mg
~ 3.9 FALGPA U/mg
1 PZ U/mg ~ 1000 Mandl or CDU U/mg
1 PZ U/mg ~ 10 HP
U/mg
