Neutral Protease (Dispase) – Research Use Only

Neutral Protease (Dispase) is a highly purified metalloprotease derived from Bacillus polymyxa, supplied as a 5 mg lyophilized powder (Catalog #B2018270). This biotechnology-grade enzyme specifically cleaves basement membrane components (particularly type IV collagen and fibronectin) while exhibiting minimal activity against other proteins, making it an excellent tool for gentle tissue dissociation and cell isolation from epithelial tissues.

Scientific Overview

Dispase (Neutral Protease) is a zinc-dependent metalloprotease that preferentially cleaves basement membrane proteins such as type IV collagen and fibronectin while leaving most other extracellular matrix components relatively intact. This selective activity allows gentle separation of epithelial layers from underlying stroma without damaging cell viability, making it a standard enzyme for primary cell isolation, epidermal sheet preparation, and tissue engineering workflows. At 36 kDa, it offers a well-characterized and reproducible tool for controlled enzymatic dissociation.

This Neutral Protease (Dispase) is suitable for:

• Gentle dissociation of epithelial tissues from stroma
• Isolation of primary cells (keratinocytes, endothelial cells, etc.)
• Preparation of intact epithelial sheets for transplantation or culture
• Studies of basement membrane integrity and remodeling
• Enzymatic treatment in tissue engineering and regenerative medicine

Usage & Handling Guidance

Store the lyophilized powder at 2–8 °C. Reconstitute in sterile PBS or HBSS to the desired concentration (typically 0.5–2.4 U/mL or 0.5–5 mg/mL depending on tissue type). Incubate tissues at 37 °C with gentle agitation for 30–120 minutes. Optimize time and concentration for each tissue to achieve the best cell yield and viability.

• Recommended applications: Tissue dissociation and epithelial sheet separation
• Working concentration: 0.5–5 mg/mL or 0.5–2.4 U/mL (optimize for specific tissue and protocol)
• Stability: Lyophilized form is stable at 2–8 °C; reconstituted solutions should be used fresh
• Handling: Use sterile techniques; gentle agitation during incubation improves dissociation

What You Get

• 5 mg Neutral Protease (Dispase) as lyophilized powder
• 36 kDa metalloprotease with high specificity for basement membrane components
• Biotechnology-grade enzyme optimized for gentle tissue dissociation
• Convenient quantity for method development and small-to-medium scale isolations
• For research use only (RUO)

Why Researchers Choose It

• Highly purified Dispase with strong activity against type IV collagen and fibronectin
• Gentle dissociation that preserves cell viability and epithelial sheet integrity
• Standard enzyme for isolating primary cells and preparing epidermal sheets
• Stable lyophilized format with reliable lot-to-lot performance
• Well-suited for tissue engineering, regenerative medicine, and cell biology research

Frequently Asked Questions (FAQ)

• What is Dispase primarily used for?
  It is used for gentle enzymatic separation of epithelial layers from underlying stroma and for primary cell isolation.
• How should I reconstitute the powder?
  Dissolve in sterile PBS or HBSS to 0.5–5 mg/mL. Use fresh working solutions for best results.
• What tissues is it best suited for?
  It works well on skin, cornea, lung, mammary gland, and other epithelial tissues.
• Is it suitable for single-cell RNA sequencing?
  Yes, when used under optimized gentle conditions to maintain high cell viability.
• How should I store the product?
  Store the lyophilized powder at 2–8 °C.

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References

• Stenn KS, Link R, Moellmann G, Madri J, Kuklinska E. Dispase, a neutral protease from Bacillus polymyxa, is a powerful fibronectinase and type IV collagenase. J Invest Dermatol. 1989 Aug;93(2):287-90.
• Nagata M, Matsumura T. Action of the bacterial neutral protease, dispase, on cultured cells and its application to fluid suspension culture with a review on biomedical application of this protease. Jpn J Exp Med. 1986 Dec;56(6):297-307.
• Umbach JA, Gundersen CB, Baker PF. Giant synaptosomes. Nature. 1984 Oct 4-10;311(5985):474-7.
• O’Gorman D, Kin T, Pawlick R, Imes S, Senior PA, Shapiro AM. Clinical islet isolation outcomes with a highly purified neutral protease for pancreas dissociation. Islets. 2013 May-Jun;5(3):111-5.
• Craviso GL. Generation of functionally competent single bovine adrenal chromaffin cells from cell aggregates using the neutral protease dispase. J Neurosci Methods. 2004 Aug 30;137(2):275-81.
• Fiebig D, Storka J, Roeder M, Meyners C, Schmelz S, Blankenfeldt W, Scrima A, Kolmar H, Fuchsbauer HL. Destructive twisting of neutral metalloproteases: the catalysis mechanism of the Dispase autolysis-inducing protein from Streptomyces mobaraensis DSM 40487. FEBS J. 2018 Nov;285(22):4246-4264.
• Fujita T, Shiota K, Yoshikawa J, Ogawa S, Aoyagi H. Simple method for analyzing the purity of protease-containing samples by acid-treatment SDS-PAGE. J Biosci Bioeng. 2019 Nov;128(5):630-635.
• Kitano Y, Okada N. Separation of the epidermal sheet by dispase. Br J Dermatol. 1983 May;108(5):555-60.
• Geggel HS, Gipson IK. Removal of viable sheets of conjunctival epithelium with dispase II. Invest Ophthalmol Vis Sci. 1985 Jan;26(1):15-22.
• Cassiman JJ, Brugmans M, Van den Berghe H. Growth and surface properties of dispase dissociated human fibroblasts. Cell Biol Int Rep. 1981 Feb;5(2):125-32.