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
Dispasa® II
protease
Sinónimos:
Proteasa from Bacillus polymyxa
Iniciar sesión para ver los precios por organización y contrato.
Seleccione un Tamaño
Cambiar Vistas
| Tamaño de envase | SKU | Disponibilidad | Precio |
|---|---|---|---|
| 1 g | Disponible para enviar HOYdesdeMILWAUKEE | $246.00 |
Acerca de este artículo
Specific activity:
≥0.5 units/mg solid
Servicio técnico
¿Necesita ayuda? Nuestro equipo de científicos experimentados está aquí para ayudarle.
Permítanos ayudarleQuality 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
La enzima de Sigma se ha utilizado en el desarrollo de un protocolo para el cultivo ex vivo de botones mamarios embrionarios de ratón.[1] Se ha utilizado en el tratamiento de fragmentos de corazón de rata durante el aislamiento de las mitocondrias del corazón de rata.[2] Se ha utilizado también para el aislamiento de células madre de la pulpa dental (DPSC) mediante hidrólisis enzimática. Estas células se han comparado con las DPSC aisladas mediante el método de explante para analizar sus propiedades de células madre y de diferenciación.[3]
La Dispase® II se ha utilizado para la clasificación de células activadas por fluorescencia (FACS).[4] Se ha utilizado también para la separación de las capas del saco vitelino visceral.[5]
Adecuada para utilizar en la preparación de suspensiones monocelulares para secuenciación.
La Dispase® II se ha utilizado para la clasificación de células activadas por fluorescencia (FACS).[4] Se ha utilizado también para la separación de las capas del saco vitelino visceral.[5]
Adecuada para utilizar en la preparación de suspensiones monocelulares para secuenciación.
Biochem/physiol Actions
Dispase®II es una proteasa neutra que hidroliza los enlaces peptídicos N-terminales de restos de aminoácidos no polares. Puede utilizarse para separar muchos tejidos y células en crecimiento in vitro. La enzima es muy suave y no daña las membranas celulares. También puede utilizarse para evitar la aglutinación en los cultivos en suspensión. Esta proteasa escinde la fibronectina y el colágeno de tipo IV, pero no la laminina, el colágeno de tipo V, la seroalbúmina ni la transferrina.[6] Dispase® II es específica para la rotura de los enlaces leucina-fenilalanina. El Ca2+, el Mg2+, el Mn2+, el Fe2+, el Fe3+ y el Al3+ activan la enzima. El EDTA, el EGTA, el Hg2+ y otros metales pesados inhiben la actividad de la enzima.[7] La enzima contiene un átomo gramo de cinc por mol gramo de enzima purificada. Si se elimina el componente cinc con quelantes como el EDTA o el EGTA, se obtiene una apoenzima inactiva. La enzima no es inhibida por el suero.
Physical form
polvo liofilizado que contiene acetato de calcio y lactosa
Preparation Note
Puede prepararse una disolución de reserva de 50 U/ml mediante la disolución del polvo en un tampón que contenga NaAc 10 mM (pH 7,5) y CaAc 5 mM. Debe conservarse a 4 °C.
Analysis Note
La enzima puede disolverse inicialmente en Hepes/KOH 50 mM pH 7,4, NaCl 150 mM a 10 mg/ml Deben realizarse más diluciones en los medios de cultivo celular más apropiados para que las células se disocien. La dispasa es una metaloproteinasa y es activada por cationes divalentes, como el Ca2+, el Mg2+, el Mn2+ y el Fe2+.
Other Notes
Una unidad hidrolizará la caseína para producir color equivalente a 1,0 μmol (181 μg) de tirosina por minuto a pH 7,5 a 37 °C (color por el reactivo Folin-Ciocalteu, salvo que se indique otra cosa).
Legal Information
Dispase is a registered trademark of Godo Shusei Co., Ltd.
1 of 1
Este artículo | |||
|---|---|---|---|
| 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 |
| 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 |
| form lyophilized solid | form lyophilized solid | form essentially salt-free, lyophilized powder | form essentially salt-free, lyophilized powder |
| 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 |
| storage temp. 2-8°C | storage temp. 2-8°C | storage temp. −20°C | storage temp. −20°C |
| 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
Clase de almacenamiento
11 - Combustible Solids
flash_point_f
Not applicable
flash_point_c
Not applicable
Elija entre una de las versiones más recientes:
¿Ya tiene este producto?
Encuentre la documentación para los productos que ha comprado recientemente en la Biblioteca de documentos.
Artículos
Use of MULTI-seq lipid-modified oligos, protocol, and troubleshooting guide for PCR Assays and Sequencing applications.
Contenido relacionado
Collagenase Guide.Collagenases, enzymes that break down the native collagen that holds animal tissues together, are made by a variety of microorganisms and by many different animal cells.
Organoid culture products to generate tissue and stem cell derived 3D brain, intestinal, gut, lung and cancer tumor organoid models.
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
Some properties of a protease from Bacillus polymyxa.
P J Griffin et al.
The Biochemical journal, 125(4), 109P-109P (1971-12-01)
Número de artículo de comercio global
| SKU | GTIN |
|---|---|
| D4693-1G | 04061833574041 |
-
Can I also dissolve the powder in 1x PBS?
1 answer-
Helpful?
-
-
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,
1 answer-
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.
Helpful?
-
-
How long can I store the powder at -80°C?
1 answer-
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.
Helpful?
-
-
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.
Helpful?
-
-
how many units in 1 mg for the product D4693
1 answer-
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?
-
-
Once dissolved into a stock solution, how long is it stable at 4 degrees C?
1 answer-
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?
-
-
Is Dispase II esteril for isolating cells from tissues for doing cell cultures? Thanks Monica
1 answer-
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?
-
-
What is the Department of Transportation shipping information for this product?
1 answer-
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.
Helpful?
-