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SCR006 Accumax Cell Counting Solution in DPBS Sterile-filtered Cell Culture Tested

A cell detachment solution of proteolytic, collagenolytic & DNAse enzymes. The reagent is useful for creating single cell suspensions from clumped cell cultures for accurate cell counting, detachment of cells from primary tissue.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
SCR006
100 mL  
Purchase on Sigma-Aldrich

Overview

Replacement Information
Description
Catalogue NumberSCR006
Brand Family Chemicon®
Trade Name
  • Chemicon
DescriptionAccumax Cell Counting Solution in DPBS Sterile-filtered Cell Culture Tested
OverviewA cell detachment solution of proteolytic, collagenolytic and DNAse enzymes. Useful on a variety of cells lines however not all cells or cell types are affected by ACCUMAX™, also passage number and confluency can influence how well the reagent does because the amounts and makeup of the collagen extracellular matrix will vary with age and culture state. Because ACCUMAX™ is a more gentle, less univerally proteolytic cleavage mixture reagent amounts and incubation times need to be optimized for each cell system.

The reagent is useful for creating single cell suspensions from clumped cell cultures for accurate cell counting, detachment of cells from primary tissue and the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware, including SmartPlastic™. ACCUMAX™ does not contain mammalian or bacterial derived products.

Each lot of ACCUMAX™ is tested for Sterility (by USP membrane filtration method), enzymatic activity (tested with synthetic chromagenic tetrapeptides) and cell detachment from tissue culture plastic.



ACCUMAX™ has been used successfully on numerous cell types including: CHO, BHK, 293, COS, Sf9, and catfish primary hepatocyte cells.
References
Product Information
HS Code3821 00 00
Presentation100 ml, ready to use, frozen sterile filtered (0.2 micron) liquid. 1X ACCUMAX™ enzymes 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 0.5mM EDTA-4Na.
Quality LevelMQ300
Applications
ApplicationA cell detachment solution of proteolytic, collagenolytic & DNAse enzymes. The reagent is useful for creating single cell suspensions from clumped cell cultures for accurate cell counting, detachment of cells from primary tissue.
Application NotesA. Cell Counting.

1. Thaw ACCUMAX™ at room temperature.

2. Harvest a representative sample of clumped cells, 0.5 or 1.0 ml, and place in the container used for cell counting.

3. Add an equal volume of ACCUMAX™ to the sample of cells, and incubate for 5 to 10 minutes at 37° C.

4. Count the cells by your normal procedure. Note that the cells have been diluted an extra 2 fold.

5. Count cells and passage as usual; no additional washes or enzyme inhibitors are required.

B. Primary Tissue Dissociation.Recent research has revealed some unique properties of the enzymes contained in ACCUMAX™. The ACCUMAX™ enzymes are more effective at room temperature than at 37°C. In fact, the ACCUMAX™ enzymes will be inactivated after two hours at 37°C. For this reason, perform all your tissue digestion at room temperature.

This protocol for using ACCUMAX™ to dissociate cells from primary tissue is a general-purpose protocol and may not be applicable to all tissue types. The individual investigator needs to optimize the conditions for his/her tissue specimens. Keep in mind that ACCUMAX™ is a powerful enzyme mixture that can potentially dissolve not only the connective tissue of solid tissue but some fragile cell types as well if not closely monitored.





Optimal incubation times and reagent strength needs to be determined for each cell/tissue type tested by the enduser.



Non-sterile materials required:

Platform rocker; Microscope; Centrifuge; Trypan Blue

Sterile Materials required:

ACCUMAX™; T25 culture flasks; DPBS (calcium and magnesium free); Centrifuge tubes (15-50 ml);

Culture medium; Scalpels; Pipettes (1 ml, 10 ml); Forceps; Petri dishes (100 mm, non-tissue culture grade)

Procedure:

1. Transfer the tissue to a petri dish containing fresh, sterile DPBS, and rinse.

2. Transfer the tissue to a second dish; dissect off unwanted tissue, such as fat or necrotic material.

3. Using two crossed scalpels or a scalpel and forceps, cut the tissue into small pieces approximately 1 mm in size.

4. Transfer the tissue pieces to a 15 or 50 ml sterile centrifuge tube containing fresh, sterile DPBS.

5. Allow the pieces to settle and carefully remove the supernatant. Repeat this wash step two times.

6. Transfer the tissue pieces to a fresh petri dish and add enough ACCUMAX™ to the plate to cover tissue.

7. Incubate the samples on a platform rocker at room temperature for 5 to 60 minutes. The tissue will “smear” on the bottom of the dish when the disaggregation is effective. To release more cells, gently agitate the sample by pipetting several times. It is best to check cell viability several times during the incubation using Trypan blue.

8. Once disaggregation is complete, transfer the cells to a sterile centrifuge tube and centrifuge at 300 x g to pellet the cells and to remove the cell debris if desired.

9. Carefully remove the supernatant and re-suspend the cell pellet in 5 ml of DMEM/F12 containing 10 – 20% FBS (or other appropriate media). Seed in a T25 flask. Replace the media after 48 hours.

For Soft Tissue samples:

1. If cell isolation is from a soft tissue (such as liver), transfer the tissue to a petri dish containing fresh, sterile DPBS, and rinse.

2. Transfer the tissue to a second dish; dissect off unwanted tissue, such as fat or necrotic material. Add 1 – 2 ml of ACCUMAX ™ and use forceps to gently “tease” the cells into the ACCUMAX™.

3. Residual connective tissue may be separated by allowing the pieces to settle or by filtration, if desired.

4. Centrifuge the sample at 300 x g to pellet the cells and to remove cell debris if desired.

5. Carefully remove the supernatant and re-suspend the cell pellet in 5 ml of DMEM/F12 containing 10 – 20% FBS (or other appropriate media). Seed in a T25 flask. Replace the media after 48 hours.
Biological Information
Media FormLiquid
Stem Cell Type
  • Mouse Embryonic Stem Cells
  • Human Embryonic Stem Cells
  • Mesenchymal Stem Cells
  • Neural Stem Cells
  • Hematopoietic Stem Cells
  • Epithelial Cells
  • Pancreatic Stem Cells
  • Cardiac Stem Cells
  • Induced Pluripotent Stem Cells
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Usage Statement
  • 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 and Shipping Information
Storage ConditionsStable when stored at -20°C. Refer to lot expiration date. Recommended storage upon receipt is -20°C. After thawing, ACCUMAX may be stored for up to 2 months at 4°C. DO NOT STORE AT ROOM TEMP. Use sterile technique when handling.
Packaging Information
Material Size100 mL
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
SCR006 04053252266652

Documentation

Accumax Cell Counting Solution in DPBS Sterile-filtered Cell Culture Tested SDS

Title

Safety Data Sheet (SDS) 

Accumax Cell Counting Solution in DPBS Sterile-filtered Cell Culture Tested Certificates of Analysis

TitleLot Number
ACCUMAX - 2080265 2080265
ACCUMAX - 2003006 2003006
ACCUMAX - 2139998 2139998
ACCUMAX™ 3007969
ACCUMAX™ 3102633
ACCUMAX™ 2943260
ACCUMAX™ 2897743
ACCUMAX™ 3077598
ACCUMAX™ 3057582
ACCUMAX™ 3028919
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Technical Info

Title
A Comparative Analysis of Human Embryonic Stem Cells Cultured in a Variety of Media Conditions
Accumax Dissociation Reagent for the Serial Passaging of Human Embryonic Stem Cells
Collagenase Type I for Enzymatic Passaging of Human Embryonic Stem Cells in HEScGRO Medium

Data Sheet

Title
ACCUMAX™
Reprogramming Cell Fate and Function Novel Strategies for iPSC Generation, Characterization, and Differentiation

Posters

Title
Poster: A Novel Serum-Free and Animal Component-Free Culture Medium for the Propagation and Maintenance of Human Embryonic Stem Cells

FAQ

QuestionAnswer
What is the difference between Accuma and Accutase?Accumax (Catalog #SCR006) will dissociate clumped cells in minutes resulting in a single cell suspension. It will yield accurate reproducible cell counts, is cost effective and is ready to use. The Accutase (Catalog #SCR005) contains phenol red. It will detect adherent cells within minutes and will dissociate tissues for primary cell culture use. It also will yield accurate reproducible cell counts, is cost effective and is ready to use.