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D4693

Sigma-Aldrich

Dispase® II

protease

Synonym(s):

Protease from Bacillus polymyxa

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About This Item

CAS Number:
42613-33-2
EC Number:
255-914-4
MDL number:
MFCD02253017
UNSPSC Code:
12352204
NACRES:
NA.54
Pricing and availability is not currently available.

Quality Level

100

form

lyophilized solid

specific activity

≥0.5 units/mg solid

technique(s)

cell culture | mammalian: suitable
single cell analysis: suitable

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.[1] It has been used in the treatment of rat heart pieces during the isolation of mitochondria from rat heart.[2] 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.[3]
Dispase® II has been used for Fluorescence-Activated Cell Sorting (FACS).[4] It has also been used for separating visceral yolk sac layers.[5]
Suitable for use in preparation of single cell suspension for sequencing.

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.[6] 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.[7] 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.

Pictograms

Health hazardExclamation mark
GHS08,GHS07

Signal Word

Danger

Hazard Statements

H315,H319,H334,H335

Hazard Classifications

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

Target Organs

Respiratory system

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

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Certificates of Analysis (COA)

Lot/Batch Number
BCCL2638
BCCM1253
BCCL7124
BCCJ7416
BCCK9179

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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
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
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
Some properties of a protease from Bacillus polymyxa.
P J Griffin et al.
The Biochemical journal, 125(4), 109P-109P (1971-12-01)
Feng-Ming Ding et al.
Molecular medicine reports, 16(1), 778-786 (2017-06-01)
Neutrophilic airway inflammation in chronic lung infections caused by Pseudomonas aeruginosa (PA) is associated with T helper (Th)17 responses. Suppressor of cytokine signaling 3 (SOCS3) is the major negative modulator of Th17 function through the suppression of signal transducer and activator
View All Related Papers

Questions

1–5 of 5 Questions  

  1. Could I dissolve in another work solution instead Hepes/KOH 50 mM pH 7,4, NaCl 150 mM? 

    1 answer

    1. Two approaches are available for reconstituting this lyophilized enzyme formulation. The first approach is for making a solution used in cell dissociation, and the second is a more generic method, such as for assaying Dispase activity:

      1. Dissolve 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.

      2. A 50 U/mL stock solution may be prepared by dissolving the powder in a buffer containing 10 mM sodium acetate (pH 7.5) and 5 mM calcium acetate.

      Helpful?

  2. how many units in 1 mg for the product D4693

    1 answer

    1. The enzymatic activity varies from lot to lot and is reported in the Certificate of Analysis. A brief survey of recent lots reveals an average activity of 1.2 u/mg solid. Please see the link below to review a sample or lot specific Certificate:
      https://www.sigmaaldrich.com/product/sigma/d4693*product-documentation

      Helpful?

  3. Once dissolved into a stock solution, how long is it stable at 4 degrees C?

    1 answer

    1. The reconstituted stock solution can be stored at 2 to 8 °C for 2 weeks. Alternatively, it can be stored at - 20 deg C in aliquots to minimize freeze-thaw cycles where it should be stable for up to 2 months. The working solution, diluted in PBS from the stock solution, is stable for approximately 3 days at 2 to 8 °C.

      Helpful?

  4. Is Dispase II esteril for isolating cells from tissues for doing cell cultures? Thanks Monica

    1 answer

    1. This product has been used in the ex vivo culture of mouse embryonic mammary buds. Please see protocols in the published article: https://link.springer.com/article/10.1007/s10911-013-9288-2

      Helpful?

  5. What is the Department of Transportation shipping information for this product?

    1 answer

    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

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