NEW! CLOSTRIPAIN, Improves islet and cell yields
Posted by Sara On Tue 07 May, 2019

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.

 


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