Pronase E (Activity ≥ 4000 U/mg) MedChemExpress (MCE)

Product Name	Pronase E
Synonyms	Proteline Pronase E EC 3.4.24.4 s.Griseusproteinase Streptomycesgriseusprotease pronasef.streptomycesgriseus Proteinase,streptomycesgriseus Synonyms Proteline Pronase E EC 3.4.24.4 s.Griseusproteinase Streptomycesgriseusprotease pronasef.streptomycesgriseus Proteinase,streptomycesgriseus ACTINASE E, STREPTOMYCES GRISEUS Protease from Streptomyces griseus,Actinase E, Pronase E
CAS	9036-06-0
EINECS	232-909-5
Chemical Formula
Molecular Weight
inchi
Package	100 mg
Price	Email to quote
Descriptions	Pronase E (Activity ≥ 4000 U/mg) Pronase E (Activity ≥ 4000 U/mg) MedChemExpress (MCE) HY-114158A 9036-06-0 Pronase (Activity ≥ 4000 U/mg) Please store the product under the recommended conditions in the Certificate of Analysis. Room temperature in continental US Descriptions Pronase E (Activity ≥ 4000 U/mg) Pronase E (Activity ≥ 4000 U/mg) MedChemExpress (MCE) HY-114158A 9036-06-0 Pronase (Activity ≥ 4000 U/mg) Please store the product under the recommended conditions in the Certificate of Analysis. Room temperature in continental US may vary elsewhere. Pronase E (Activity ≥ 4000 U/mg) is a mixture of proteolytic enzymes that is obtained from Streptomyces griseus and could digest protein into individual amino acids. Pronase E significantly increases the amount of most of the amino acids analysed (PE vs C) , especially Ile, His and Thr[1]. Pronase E Usage Steps:. 1. Preparation of Stock Solution: (1) Dissolve Pronase E in deionized water to prepare a stock solution at a concentration of 5-20 mg/ml. Note: If used for DNA or RNA isolation steps, it is recommended to first heat the solution at 56°C for 15 minutes. (2) Incubate at 37°C for 1 hour. Aliquot the prepared stock solution into single-use samples and store at -20°C typically stable for one year. 2. DNA Isolation: Directly add Pronase E to the DNA preparation system. The system should contain 0.5-1% SDS to disrupt DNA-protein interactions. Note: Use at a concentration of 250-500 μg protein/ml. Incubation conditions: Incubate at 37°C for 1-4 hours. 3. Protein Digestion: (1) Dissolve approximately 0.2 μM protein in 0.2 ml of 50 mM ammonium bicarbonate buffer (pH 8.0) or phosphate buffer (pH 7.0). (2) Add 1% (w/w) Pronase E. Incubate at 37°C for 24 hours to perform protein digestion. 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. \| \| \| \| \| \| \| \| \| \| [1]. José M Bruna, et al. Enhancement of the flavour development of dry fermented sausages by using a protease (Pronase E) and a cell‐free extract of Penicillium camemberti. Journal of the science of food and agriculture. Volume82, Issue5. April 2002. Pages 526-533. [2]. Chen QT, et al. HK1 from hepatic stellate cell-derived extracellular vesicles promotes progression of hepatocellular carcinoma. Nat Metab. 2022 Oct 4(10):1306-1321. [Content Brief]
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Last Update	2025-04-01 14:45:47

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