- Pronase E
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- $0.00 / 1g
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2025-04-16
- CAS:9036-06-0
- Min. Order: 1g
- Purity: 98.0%
- Supply Ability: 1kg/month
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| | EC 3.4.24.4 Basic information |
| Product Name: | EC 3.4.24.4 | | Synonyms: | s.Griseusproteinase;Streptomycesgriseusprotease;Streptomycesgriseusproteinase;ACTINASE E, STREPTOMYCES GRISEUS;Protease from Streptomyces griseus,Actinase E, Pronase E;pronasef.streptomycesgriseus;Proteinase,streptomycesgriseus;Proteline | | CAS: | 9036-06-0 | | MF: | NULL | | MW: | 0 | | EINECS: | 232-909-5 | | Product Categories: | enzyme | | Mol File: | Mol File | ![EC 3.4.24.4 Structure]() |
| | EC 3.4.24.4 Chemical Properties |
| storage temp. | 2-8°C | | solubility | H2O: 5-20 mg/mL | | form | powder | | color | slightly brown | | biological source | Streptomyces griseus | | Water Solubility | water: 10mg/mL | | Specific Activity | ≥70,000proteolytic units/g dry wt |
| | EC 3.4.24.4 Usage And Synthesis |
| Uses | Pronase E can be used to degrade antheraea pernyi silk fibroin films. | | General Description | Note: 1 KU = 1000 units. | | Biochem/physiol Actions | This product is a mixture of at least three caseinolytic activities and one aminopeptidase activity. The caseinolytic enzymes were named as Streptomyces griseus Protease A, Streptomyces griseus Protease B and Streptomyces griseus Trypsin. This product may be used when extensive or complete degradation of protein is required. This protease mixture is highly nonspecific and can digest casein to the extent of >70% as mono-amino acids. | | in vivo | Experimental Procedure for Isolating Primary Hematopoietic Stem Cells (HSCs) from Mice Using Pronase E[2]1. Animal Anesthesia and Abdominal Exposure:
(1) Anesthetize male or female mice.
(2) Under sterile conditions, expose the liver via abdominal incision.
2. Liver Perfusion Procedure:
(1) Perform liver perfusion through the portal vein using Hanks' Balanced Salt Solution (HBSS) to remove blood and impurities.
(2) Conduct enzymatic perfusion with Pronase E and collagenase to dissociate liver tissue.
3. Liver Extraction and Cell Release:
(1) Carefully extract the liver from the abdominal cavity using sterile forceps, and quickly transfer it to a sterile culture dish.
(2) Gently peel the liver capsule and apply mild mechanical agitation to release cells from the liver tissue.
4. Cell Digestion and Preliminary Centrifugation:
(1) Suspend the released cells in a mixed enzyme solution containing Pronase E, collagenase, and DNase, and digest at 37°C for 20 minutes.
(2) Perform two rounds of low-speed centrifugation (50 g, 3 minutes) to separate liver cells, and collect the cell suspension.
5. HSC Separation and Density Gradient Centrifugation:
(1) Further centrifuge the cell suspension (450 g, 8 minutes) and collect the pellet.
(2) Resuspend the pellet in 18% Nycodenz solution as the basis for density gradient separation.
(3) Construct a density gradient with 12% Nycodenz, 8.2% Nycodenz, and Gey's Balanced Salt Solution (GBSS).
(4) Isolate and purify HSCs from the 8.2% Nycodenz layer through density gradient centrifugation.
Note: This protocol provides standard operating procedures; please adjust and optimize according to specific experimental needs and conditions. |
| | EC 3.4.24.4 Preparation Products And Raw materials |
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