D4693

Sigma-Aldrich

Dispase® II

protease

Synonym(s):
Protease from Bacillus polymyxa
CAS Number:
EC Number:
MDL number:
NACRES:
NA.54

Quality Level

form

lyophilized solid

specific activity

≥0.5 units/mg solid

solubility

10 mM NaAc (pH 7.5) and 5 mM CaAc: soluble

storage temp.

2-8°C

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Application

The enzyme from Sigma has been used in developing a protocol for ex vivo culture of mouse embryonic mammary buds. It has been used in the treatment of rat heart pieces during the isolation of mitochondria from rat heart. It has also been used for the isolation of dental pulp stem cells (DPSCs) by enzymatic hydrolysis. These cells have been compared with DPSCs isolated by explant method to analyse their stem cell and differentiation properties.
Dispase® II has been used for Fluorescence-Activated Cell Sorting (FACS). It has also been used for separating visceral yolk sac layers.

Biochem/physiol Actions

Dispase® II is a neutral protease that hydrolyzes the N-terminal peptide bonds of non-polar amino acid residues. It may be used for separating many tissues and cells grown in vitro. The enzyme is very gentle and does not damage cell membranes. It can also be used to prevent clumping in suspension cultures. This protease cleaves fibronectin and type IV collagen, but not laminin, type V collagen, serum albumin, or transferrin. Dispase® II is specific for the cleavage of Leucine-Phenylalanine bonds. Ca2+, Mg2+, Mn2+, Fe2+, Fe3+ and Al3+ activate the enzyme. EDTA, EGTA, Hg2+ and other heavy metals inhibit the enzyme activity. The enzyme contains 1g-atom of zinc per g-mol of purified enzyme. If this zinc component is removed by chelating agents such as EDTA or EGTA, an inactive apoenzyme is obtained. The enzyme is not inhibited by serum.

Unit Definition

One unit will hydrolyze casein to produce color equivalent to 1.0 μmole (181 μg) of tyrosine per min at pH 7.5 at 37 °C (color by Folin-Ciocalteu reagent), unless otherwise indicated.

Physical form

lyophilized powder containing calcium acetate and milk sugar

Preparation Note

A 50 U/mL stock solution may be prepared by dissolving the powder in a buffer containing 10 mM NaAc (pH 7.5) and 5 mM CaAc. It should be stored at 4 °C.

Analysis Note

The enzyme can be initially dissolved in 50 mM Hepes/KOH pH 7.4, 150 mM NaCl at 10 mg/ml. Further dilutions should be made in the cell culture media most appropriate for the cells being dissociated. Dispase is a metalloproteinase and is activated by divalent cations including Ca2+ , Mg2+, Mn2+ and Fe2+.

Legal Information

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

RIDADR

NONH for all modes of transport

WGK Germany

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Certificate of Origin

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P Hilkens et al.
Cell and tissue research, 353(1), 65-78 (2013-05-30)
Dental pulp stem cells (DPSCs) are an attractive alternative mesenchymal stem cell (MSC) source because of their isolation simplicity compared with the more invasive methods associated with harvesting other MSC sources. However, the isolation method to be favored for obtaining...
Maria Voutilainen et al.
Journal of mammary gland biology and neoplasia, 18(2), 239-245 (2013-05-16)
The explant culture techniques of embryonic tissues allow continuous monitoring of organ growth and morphogenesis ex vivo. The effect of growth factors and other soluble molecules can be examined by applying them to the culture medium. Relatively few studies have...
Shuya Liu et al.
Frontiers in physiology, 8, 849-849 (2017-11-23)
The mouse bitter taste receptors Tas2r143, Tas2r135, and Tas2r126 are encoded by genes that cluster on chromosome 6 and have been suggested to be expressed under common regulatory elements. Previous studies indicated that the
Magdalena Labieniec-Watala et al.
International journal of molecular sciences, 12(11), 8013-8026 (2011-12-17)
Diabetes is associated with a mitochondrial dysfunction. Hyperglycaemia is also clearly recognized as the primary culprit in the pathogenesis of cardiac complications. In response to glycation and oxidative stress, cardiac mitochondria undergo cumulative alterations, often leading to heart deterioration. There...
Quanah J Hudson et al.
Developmental biology, 353(2), 420-431 (2011-03-01)
A subset of imprinted genes in the mouse have been reported to show imprinted expression that is restricted to the placenta, a short-lived extra-embryonic organ. Notably, these so-called "placental-specific" imprinted genes are expressed from both parental alleles in embryo and...
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