MilliporeSigma
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A6964

Sigma-Aldrich

Accutase® solution

sterile-filtered, suitable for cell culture

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Synonym(s):
Cell detachment solution
MDL number:
MFCD01632905
NACRES:
NA.75

Quality Level

200

sterility

sterile-filtered

form

liquid

concentration

400-600 units/mL

technique(s)

cell culture | mammalian: suitable

pH

6.8-7.8

shipped in

dry ice

storage temp.

−20°C

General description

Accutase® solution (in dulbecco′s phosphate-buffered saline (DPBS) without Ca++ or Mg++) contains proteolytic and collagenolytic enzymes for cell detachment. It is useful for the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware.

Application

Accutase® solution has been used for dissociation or digestion of nonadherent cells growing in an aggregated cluster, such as brain tumor cells, induced pluripotent stem cells (IPSCs) and glioblastoma cells into single cells.
Proven effective in detaching primary fibroblasts, endothelial cells, neurons, tumor cell lines, and insect cells. Performs exceptionally well in detaching cells for analysis of cell surface markers, virus growth assay, and flow cytometry as well as bioreactor scale-up. Does not contain mammalian or bacterial-derived products.

Features and Benefits

Ready-to-use liquid consisting of a proprietary mixture of proteolytic and collagenolytic enzymes for use in dissociation of cells from standard and adhesion-coated plasticware.

Formula variant

Prepared in Dulbecco′s PBS (0.2 g/L KCl, 0.2 g/L KH2PO4, 8 g/L NaCl, and 1.15 g/L Na2HPO4) containing 0.5 mM EDTA•4Na and 3 mg/L Phenol Red. Refer to Product Data Sheet on the website for product usage information.

Preparation Note

This product does contain phenol red. The product is shipped on dry ice and there could be significant CO2 buildup in the package. This CO2 may enter the solution and lower the pH slightly, giving an yellow-orange (around pH 6.8) vs. pinkish (around pH 7.8) color. The solution, if yellow-orange (acidic) should still be good to use as is, or sodium hydroxide may be added to adjust the pH.

Legal Information

Accutase is a registered trademark of Innovative Cell Technologies, Inc.

Storage Class Code

10 - Combustible liquids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Enter Lot Number to search for Certificate of Analysis (COA).

Certificate of Origin

Enter Lot Number to search for Certificate of Origin (COO).

Quotes and Ordering

Bulk Quote-Order Product

  1. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  2. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  3. For what purposes can Product A6964, Accutase® solution, be used?

    It can be used for detaching cells, analysis of cell surface markers and virus growth assays.

  4. Will Product A6964, Accutase® solution, work on tissue?

    This product is not tested for tissue, but it should work on soft tissue.

  5. What is the concentration of Product A6964, Accutase® solution?

    This product is typically at a concentration of 400-600 units/mL.

  6. How do I find price and availability?

    There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers.

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

    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. 

  8. Does Product A6964, Accutase® solution, contain animal components?

    Accutase is purified from a marine invertebrate.  There are no other animal or bacterial derived proteins present.

  9. What is the difference between Accutase and Accumax?

    Accutase is a mixture of non-mammalian proteases.  Accumax is accutase that has been augmented with DNase to reduce clumping associated with DNA release from damaged cells.

  10. What is the appropriate cell dissociating product to be used for ES cells that will be injected into blastocysts?

    Accutase®, product A6964, or Accumax™ (Accutase® with DNase), product A7089, will work well for dissociation of ES cells. The Accutase® product has protease and dispase activity to dissociate the cells.  The Accumax™ product also has DNase to limit the aggregation that is caused by DNA released from ruptured cells during the dissociation/scraping.Please see the reference and link below for more information on the use of Accutase® with ES cells:Mol Reprod Dev. 2008 May;75(5):818-27

  11. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

  12. How can I inactivate Accutase® with fetal bovine serum?

    Due to its gentle nature Accutase® cell detachment solution usually does not need to be neutralized following cell passaging. Accutase® is also heat inactivated after 45 minutes at 37°C. If this method of neutralization does not work for your cells, or if you want to neutralize Accutase® in a shorter period of time, the addition of 10% FBS containing media will accomplish this task. Experiments have shown that diluting a sample of Accutase® with 10% with fetal bovine serum achieved an environment similar to that found in cell culture. Measurements of the enzyme activity in this diluted sample showed decreased activity over time, falling below that of the required level for cell detachment (>175 units/mL). After about 6 minutes of serum interaction, the enzyme activity of Accutase® was well below this level, demonstrating effective inhibition by FBS.

Ruchi Bajpai et al.
Molecular reproduction and development, 75(5), 818-827 (2007-12-25)
Human embryonic stem cells (hESCs) hold great promise for cell-based therapies and drug screening applications. However, growing and processing large quantities of undifferentiated hESCs is a challenging task. Conventionally, hESCs are passaged as clusters, which can limit their growth efficiency
Samia Rourou et al.
Vaccine, 25(19), 3879-3889 (2007-02-20)
Rabies virus strain production in Vero cells grown on Cytodex 1 in a 2 L stirred tank bioreactor and in a medium free of components of human or animal origin (VP-SFM) is described. Cell banking procedure in VP-SFM supplemented with
Recent Zika virus isolates induce premature differentiation of neural progenitors in human brain organoids
Gabriel E, et al.
Cell Stem Cell, 20(3), 397-406 (2017)
Yanting Xue et al.
Stem cell research, 24, 151-154 (2017-10-17)
Human MSX1 gene is mapped to chromosome 4 and encodes a 303aa homeobox protein MSX1. MSX1 expression appears during early tooth development of vertebrate embryogenesis. Mutations in this protein are related to human tooth anomalie, cleft lip and palate and
Autophagy pathways in glioblastoma
Jiang H, et al.
Methods in Enzymology, 453, 273-286 (2009)
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