C0773

Colagenase

high purity, purified by chromatography, Type VII, ≥4 FALGPA units/mg solid, lyophilized powder, ≥700 CDU/mg solid (CDU = collagen digestion units)

• Sinônimo(s): Clostridiopeptidase A
• Número CAS: 9001-12-1
• UNSPSC Code: 12352204
• NACRES: NA.54
• EC Number: 232-582-9
• MDL number: MFCD00130830
• Número CE: 3.4.24.3 (BRENDA, IUBMB)
• Specific activity: ≥4 FALGPA units/mg solid, ≥700 CDU/mg solid (CDU = collagen digestion units)

Propriedades

• Quality Segment: 200
• type: Type VII
• form: lyophilized powder
• specific activity: ≥4 FALGPA units/mg solid, ≥700 CDU/mg solid (CDU = collagen digestion units)
• mol wt: 68-130 kDa
• purified by: chromatography
• composition: Protein, ≥85% biuret
• solubility: TESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4): soluble 0.05-0.1 mg/mL at 37 °C
• foreign activity: caseinase and clostripain ≤1 units/mg protein
• storage temp.: −20°C

Descrição

Application

Collagenase has been used in the preparation of arterial tissue for the study of advanced glycosylation end products (AGE). The enzyme has also been used along with other proteases for the disaggregation of human tumor, mouse kidney, human brain, lung epithelium and many other tissues. It is also effective in liver and kidney perfusion studies, digestion of pancreas, and isolation of nonparenchymal hepatocytes.

Biochem/physiol Actions

A colagenase é ativada por quatro gramas de átomos de cálcio por mol de enzima. Ela é inibida por ácido etileno glicol-bis(beta-aminoetil éter)-N, N, N′,N′-tetracético, beta-mercaptoetanol, glutationa, ácido tioglicólico e 8-hidroxiquinolina.

Effective release of cells from tissue requires the action of both collagenase enzymes and the neutral protease. Collagenase is activated by four gram atom calcium (Ca²⁺) per mole enzyme. The culture filtrate is thought to contain at least 7 different proteases ranging in molecular weight from 68-130 kDa. Optimal pH ranges from 6.3-8.8. The enzyme recognizes the sequence -R-Pro-8-X-Gly-Pro-R-, where X is most often a neutral amino acid.

Physical form

Lyophilized powder containing calcium chloride

Preparation Note

A stock solution may be prepared by dissolving 0.05 - 0.1 mg of collagenase in one mL of 50 mM TES buffer containing 0.36 mM calcium chloride (TESCA), pH 7.4 at 37 °C. Use cold buffer.

Please note the material will not dissolve completely and few sediments may be seen. The sediments do not affect the activity of the solution.

Other Notes

Uma unidade de digestão de colágeno (CDU) libera peptídeos de colágeno do tendão de Aquiles bovino que equivalem em cor de ninidrina a 1,0 μmol de leucina em 5 horas em pH 7,4 a 37 °C na presença de íons de cálcio. Uma unidade de hidrólise de FALGPA hidrolisa 1,0 μmol de furilacriloil-Leu-Gli-Pro-Ala por min a 25 °C. Uma unidade de protease neutra hidrolisa caseína para produzir cor equivalente a 1,0 μmol de tirosina por 5 h em pH 7,5 a 37 °C. Uma unidade de clostripaína hidrolisa 1,0 μmol de BAEE por min em pH 7,6 a 25 °C na presença de DTT.

Informações de segurança

• pictograms: GHS08,GHS07
• signalword: Danger
• hcodes: H315,H319,H334,H335
• pcodes: P261 - P264 - P271 - P280 - P302 + P352 - P305 + P351 + P338
• Hazard Classifications: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• target_organs: Respiratory system
• Classe de armazenamento: 11 - Combustible Solids
• wgk: WGK 1
• flash_point_f: Not applicable
• flash_point_c: Not applicable
• ppe: dust mask type N95 (US), Eyeshields, Faceshields, Gloves