This material may be reconstituted in 1X PBS without Calcium and Magnesium. However, when this product is prepared as recommended, in 50 mM HEPES/KOH (pH 7.4) with 150 mM NaCl to 10 mg/mL, or in 10 mM sodium acetate (pH 7.5) with 5 mM calcium acetate, solutions are expected to be stable for 2 weeks at 2-8°C and up to 2 months when stored in aliquots at -20°C. Solution stability in PBS has not been evaluated. Various sources indicate solutions diluted in PBS are stable for approximately 3 days at 2–8 °C.
D4693
Dispase® II
protease
Synonym(s):
Protease from Bacillus polymyxa
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About This Item
Specific activity:
≥0.5 units/mg solid
Pricing and availability is not currently available.
Quality Level
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
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.
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.
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+.
Other Notes
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.
Legal Information
Dispase is a registered trademark of Godo Shusei Co., Ltd.
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This Item | |||
|---|---|---|---|
| specific activity ≥0.5 units/mg solid | specific activity ≥10 unit/mg solid | specific activity ≥9,000 BAEE units/mg protein | specific activity ≥125 CDU/mg solid, 0.5-5.0 FALGPA units/mg solid |
| technique(s) cell culture | mammalian: suitable, single cell analysis: suitable | technique(s) cell culture | mammalian: suitable, single cell analysis: suitable | technique(s) cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable, single cell analysis: suitable |
| form lyophilized solid | form lyophilized solid | form essentially salt-free, lyophilized powder | form essentially salt-free, lyophilized powder |
| storage temp. 2-8°C | storage temp. 2-8°C | storage temp. −20°C | storage temp. −20°C |
| solubility 10 mM NaAc (pH 7.5) and 5 mM CaAc: soluble | solubility - | solubility hydrochloric acid: soluble 1 mM | solubility TESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4): soluble |
| Quality Level 100 | Quality Level 200 | Quality Level 200 | Quality Level 200 |
signalword
Danger
hcodes
Hazard Classifications
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
target_organs
Respiratory system
Storage Class
11 - Combustible Solids
flash_point_f
Not applicable
flash_point_c
Not applicable
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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
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
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
Global Trade Item Number
| SKU | GTIN |
|---|---|
| D4693-1G | 04061833574041 |
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Can I also dissolve the powder in 1x PBS?
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Can I aliquot in 10 mM sodium acetate (pH 7.5) + 5 mM calcium acetate, freeze once, and thaw as needed? I need small amounts from the same lot over several months, therefore I cannot store it at 2-8°degrees for only 2 weeks,
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Aliquots of this product prepared in 10 mM sodium acetate (pH 7.5) + 5 mM calcium acetate can be stored at -20 °C, where stability is known for up to 2 months.
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How long can I store the powder at -80°C?
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In powder form, this product is quite stable and stored at 2-8°C. This product is not assigned an expiration or retest date. A warranty period of 1 year from the date of shipment applies. The long term stability of this material when stored at -20°C or -80°C has not been investigated. Prepared solutions are expected to be stable for 2 weeks at 2-8°C and up to 2 months when stored in aliquots at -20°C.
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Could I dissolve in another work solution instead Hepes/KOH 50 mM pH 7,4, NaCl 150 mM?
1 answer-
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.
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how many units in 1 mg for the product D4693
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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-documentationHelpful?
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Once dissolved into a stock solution, how long is it stable at 4 degrees C?
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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.
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Is Dispase II esteril for isolating cells from tissues for doing cell cultures? Thanks Monica
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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
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What is the Department of Transportation shipping information for this product?
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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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