NEW! Clostripain
NOW AVAILABLE
Collagenase and Clostripain NB
fine tune your isolation
PRODUCT INFORMATION
Clostripain NB Cat. No.S2020501
General
Clostripain from Clostridium histolyticum is a proteinase that cleaves
peptide bonds on the carboxyl side of arginine. It is also known as
Endoproteinase Arg-C. As a proteolytic enzyme it can be used for
isolation of various cell types by tissue dissociation.
Clostripain exists in two different forms, anoxidized, native condition
and a reduced activated form that differ in enzymatic activity.
Description
Clostripain NB is chromatographically highly purified and largely free
from additional enzymatic activities.
Specification
Clostripain, native state activity ≥ 5.0 U/mg (BAEE)
Clostripain, activated activity ≥ 50.0 U/mg (BAEE)
Clostripain activity, native state, per vial ≥ 200U/vial (BAEE)
Clostripain activity, activated, per vial status
Endotoxin status
Application
Clostripain NB is used in combination with Collagenase NB 1
Premium Grade and/or Neutral Protease NB,
Collagenase NB 4 Standard Grade and NB 8
Broad Range suitable for cell isolation from various tissue
types.
Storage conditions
Clostripain NB is provided as a lyophilized powder. It should be
stored at -20 °C in a dry environment. Under these conditions the
product is stable until the minimum shelf life stated on the certificate
of analysis if repeated opening and closing of the vial is avoided.
For storage of solutions please refer to “Stock solution”.
Documents
For each lot a specific certificate of analysis is provided. A certificate
of origin is available.
PRODUCT INFORMATION
Clostripain NB
Instructions for use:
General
Clostripain NB is, in combination with Collagenase NB1 Premium
Grade and Neutral Protease NB, especially suited for isolation of
islets of Langerhans from human pancreas.
Clostripain NB can also be used in combination with Collagenase NB 4
Standard Grade and NB 8 Broad Range Grade for cell isolation from
several other human or animal tissues.
The native clostripain can be activated with reducing agents (e.g. DTT
Cysteine) resulting in a higher activity of the enzyme.
Tissue dissociation
Recommended starting concentrations for isolation of islets of
Langerhans from human pancreas:
Collagenase NB 1 Premium Grade: 15 – 20 PZ U/g tissue
Neutral Protease NB: 0.8 – 1.5 DMC U/g tissue
Clostripain NB, native state: 0.8 – 1.8 U/g tissue
Collagenase may contain traces of clostripain which have to be
considered to calculate the final clostripain concentration.
Clostripain NB is provided in vials with not less than 200 U which is
usually sufficient for dissociation of one human pancreas.
In general, the appropriate clostripain concentration depends on
tissue type and origin as well as on the isolation procedure.
Stock solution
A stock solution of Clostripain NB is prepared by dissolving the
enzyme in buffer. The enzyme solution must be constantly stored on ice.
Since Clostripain depends on calcium, absolutely no calcium
chelating agents (e.g. EDTA) should be present at all.
For 0.22 μm filtration filters with low protein-binding properties
(e.g. cellulose acetate, PVDF, or PES) are recommended.
Working solution
To prepare a working solution, the stock solution is diluted with
buffer to achieve the required clostripain concentration. The working
solution must be constantly stored on ice until use.
If Clostripain NB solution is mixed with Collagenase NB and Neutral
Protease NB solution, the blend should be used immediately.
Inactivation and inhibitors
The dissociation process can be reduced, e.g. by cooling down
or dilution of the enzyme solution.
Inhibitors of Clostripain include chelating agents like EDTA and all
oxidizing agents.
Important note
Clostripain NB is not intended for direct application in humans.