COLLDISP-RO

Collagenase/Dispase^®

lyophilized, non-sterile, optimum pH 7.0-8.0

• Synonyma: collagenase
• UNSPSC Code: 12352204

Vlastnosti

• biological source: enzyme from Bacillus polymyxa ( Dispase ); enzyme from bacterial ( Collagenase from Vibrio alginolyticus)
• Quality Level: 100
• sterility: non-sterile
• form: lyophilized
• collagenase activity: >0.1 U/mg lyophilizate, 25 °C (4-phenyl-azobenzyl-oxycarbonyl-Pro-Leu-Gly-Pro-D-Arg as substrate)
• specific activity: >0.8 U/mg lyophilizate, pH 7.5, 37 °C (casein as substrate, Dispase)
• packaging: pkg of 100 mg (10269638001); pkg of 500 mg (11097113001)
• manufacturer/tradename: Roche
• concentration: 1 mg/mL (Collagenase: 0.1U/mL, Dispase: 0.8U/mL)
• technique(s): tissue processing: suitable
• color: light yellow to brown
• optimum pH: 7.0-8.0
• solubility: water: soluble
• UniProt accession no.: P43154
• application(s): life science and biopharma; sample preparation
• foreign activity: proteolytic activities <0.1 U/mg (hemoglobin as the substrate)
• storage temp.: 2-8°C

Popis

General description

Dispase is a neutral, rapid-acting, gentle protease that separates intact epidermis from dermis. It also separates cultured intact epithelial cells from the substratum. It cleaves the basement membrane zone and leaves the epithelial cells intact.[1] Collagenase is a protease which cleaves the triple-helical protein called collagen. There are three types of tissue collagenases, and these belong to the matrix metalloproteinases (MMP) family.[2] Collagenase obtained from Clostridium histolyticum has a very strong activity, as it digests collagen from both ends, at temperatures as low as 4-10 °C.[3]

Specificity

Collagenase degrades native collagen. Dispase^® is a nonspecific protease.

Application

Collagenase/Dispase^® is used for the preparation of cells from a wide variety of different tissues and organs.[4][5][6]

Components

EC 3.4.24.4 & 3.4.24.3

Unit Definition

One unit is the enzyme activity, which liberates under assay conditions within 1 minute Folin-positive amino acids and peptides corresponding to 1 μmol tyrosine.

Preparation Note

Activator: Ca²⁺

Working concentration: Approximately 1mg/mL (Collagenase: 0.1U/mL, Dispase: 0.8U/mL)

Enzyme activity:
Collagenase: 0.1 Wünsch U/mg lyophilizate (+25°C, 4-phenyl-azobenzyl-oxycarbonyl-Pro-Leu-Gly-Pro-D-Arg as the substrate)
Dispase^®: 0.8 Wünsch U/mg lyophilizate (+37°C, casein as substrate, pH 7.5)
Contaminating proteolytic activities: <0.1U/mg lyophilizate (hemoglobin as the substrate)

Working solution: Preparation of working solution Dissolve enzyme in water to a final concentration of 100mg/mL. Dilute with PBS (Ca²⁺/Mg²⁺-free) to a final concentration of 1mg/mL and filter through a sterile 0.2μm membrane filter.
Enzyme concentration of the sterile solution: Collagenase, 0.1U/mL PBS; Dispase^®, 0.8U/mL PBS.

Inhibitors:
Collagenase inhibitors: EDTA, EGTA
Collagenase/Dispase is not inhibited by serum.

Storage conditions (working solution): -15 to -25°C
The reconstituted solution is stable at -15 to -25°C.
Note: Store in appropriate aliquots and avoid repeated freezing and thawing!

Reconstitution

Reconstitution in water (100mg/mL). Further dilution with PBS (phosphate buffered saline) (Ca²⁺/Mg²⁺-free).

Storage and Stability

Store dry

Other Notes

Collagenase from Vibrio alginolyticus, Dispase^® from Bacillus polymyxa

For life science research only. Not for use in diagnostic procedures.

Legal Information

Dispase is a registered trademark of Godo Shusei Co., Ltd.

Bezpečnostní informace

• pictograms: GHS07,GHS08
• signalword: Danger
• hcodes: H315 - H319 - H334 - H335
• pcodes: P261 - P264 - P280 - P284 - P304 + P340 + P312 - P342 + P311
• Hazard Classifications: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• target_organs: Respiratory system
• Storage Class: 11 - Combustible Solids
• wgk_germany: WGK 1
• flash_point_f: does not flash
• flash_point_c: does not flash