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SCM133

Sigma-Aldrich

PluriSTEM® Dispase-II Solution

Dispase-II has proven to be a rapid and effective, yet gentle, agent for separating many tissues and cells grown in vitro.

Synonym(s):

Dispase, Dispase 2

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About This Item

UNSPSC Code:
12352204
eCl@ss:
32160801
NACRES:
NA.75

Quality Level

form

liquid

manufacturer/tradename

PluriSTEM®

concentration

1 mg/mL in sterile DMEM/F-12

technique(s)

cell culture | mammalian: suitable
cell culture | stem cell: suitable

shipped in

dry ice

General description

Proteolytic enzymes such as trypsin, collagenase and pronase are commonly used for dispersing tissues and cells. These enzymes, however, often injure the cells, are unstable during incubation, can be heterogeneous and also a source of mycoplasma contamination. Dispase-II is used for the preparation of cells from a wide variety of different tissues and organs. Dispase-II has proven to be a rapid and effective, yet gentle, agent for separating many tissues and cells grown in vitro. Dispase is especially suitable for tissue disaggregation and subcultivation procedures since it does not damage cell membranes.

PluriSTEM Dispase-II has been validated to work alongside PluriSTEM Human ES/iPS Medium (SCM130) for the culture and passage of human embryonic and induced pluripotent stem cells.

Dispase-II enzyme is produced in Bacillus polymyxa.

Application

Optimal passaging technique must be determined by end user and varies depending on cell type and culture conditions.

1. Coat 6-well plates with 1:20 Matrigel coating (1.5 ml per well). Incubate at 2-8ºC overnight or coat at RT for at least 30 minutes to 1 hour before use.
2. Remove matrigel coating. Add 2 mL of Complete Media per well.
3. Remove areas of differentiation within culture.
4. Aspirate media.
5. Wash once with 2 mL PBS.
6. Add 1.5 mL Dispase II per well. Incubate for 37ºC for 7 minutes.
7. Wash 2X with 2 mL PBS, w/o Mg and Ca.
8. Add 2 mL Complete Media to each well and use cell scraper to detach colonies.
9. Collect scrapped cells in 15 ml conical tube.
10. Spin 800 rpm for 5 minutes
11. Resuspend in appropriate volume of complete media. Typical splitting ratio is 1:5 – 1:6 depending upon cell density. Cells should be ready for passaging in 5-6 days time using PluriSTEM Human ES/iPS Medium.
Research Category
Stem Cell Research
Research Sub Category
Enzymes & Biochemistry

Quality

Solution is red with no particulates present. Solution is sterile and suitable for cell culture applications.

Physical form

Product is presented in sterile DMEMF12 at 1 mg/ml. Product is filtered through a 0.2 micron filter before freezing.
Product is presented in sterile DMEMF12 at 1 mg/ml. Product is filtered through a 0.2 micron filter before freezing.
Dispase-II enzyme is produced in Bacillus polymyxa.

Storage and Stability

Store at -20°C for up to 4 months from date of receipt. Once thawed, aliquot in smaller working volumes and store at -20ºC. Avoid multiple freeze thaw cycles to maintain proper enzymatic activity.

Legal Information

PLURISTEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage Class Code

10 - Combustible liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Articles

Skip weekend feedings. Defined serum-free and feeder-free expansion media for human pluripotent stem cells (ES and iPS cells). See publications and protocols.

Skip weekend feedings. Defined serum-free and feeder-free expansion media for human pluripotent stem cells (ES and iPS cells). See publications and protocols.

Skip weekend feedings. Defined serum-free and feeder-free expansion media for human pluripotent stem cells (ES and iPS cells). See publications and protocols.

Skip weekend feedings. Defined serum-free and feeder-free expansion media for human pluripotent stem cells (ES and iPS cells). See publications and protocols.

See All

Protocols

Stem cell reprogramming protocols to generate human induced pluripotent stem cells (iPSCs) including viral and non-viral RNA based methods.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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