Accutase Solution

Accutase is an enzyme-containing cell detachment solution for adherent cells and represents a gentle alternative to conventional trypsin treatment. Its unique formulation of proteolytic and collagenolytic enzymes enables gentle and effective cell detachment from standard cell culture vessels and adhesion-coated plasticware. Compared to cell detachment with trypsin/EDTA, our Accutase is characterized by its easier handling (inactivation of the enzymes by inhibitors or serum is not necessary. Simply dilute the cell suspension to stop detachment) and higher cell viability due to reduced damage to the cells.

Our Accutase is free of mammalian or bacterial components and shows good detachment effects on adherent human and animal cells as well as insect cells. It can be used for routine detachment of cells for cell passaging or analysis of cell surface markers, cell proliferation and cell haptotaxis as well as for flow cytometry, virus growth assays, transformation assays by oncogenic transfection, neural crest cell migration assays, tumor cell migration assays, production scaling (bioreactor).

Accutase without Phenol Red

There is evidence that phenol red may interfere with hormonal activity. For this reason, we also offer an Accutase solution without phenol red, which is particularly suitable for research on endocrine cells or for studying hormonal cycles and their influence on metabolism, growth, and reproduction. Accutase retains its gentle cell-dissociation properties.

Accutase without phenol red is available as a 3x stock solution (33.3 mL) together with 500 mL of PBS without Ca²⁺ and Mg²⁺. Before use, please dilute the Accutase stock solution 3:1 (to obtain a final 100 mL solution at 1x concentration) with PBS without Ca²⁺ and Mg²⁺.

High cell viability - Gentle and effective cell detachment
Easy handling - No inactivation necessary
Composition - With EDTA, with/without phenol red

Cultured cells must be regularly passaged (split, subcultured) so that they are not exposed to too close cell-cell contacts and reduce their cell division rate by inhibiting cell contact. The passaging rate depends on the cell type (standard culture conditions: 37°C, 5% CO2 at a confluence of approx. 80%). In order to passagate adherent cells or prepare them for analysis, they must first be detached from their culture vessels.

The most common method is the so-called trypsinization, a cell separation using trypsin or trypsin/EDTA solutions. Trypsin is a serine protease and cleaves peptide bonds on the C-terminal side of basic amino acids such as lysine and arginine. During trypsinization, extracellular adhesion proteins, through which the cells adhere to the surface of culture vessels, are cleaved. However, the longer trypsin can act on the cells, the more proteins are cleaved and important cellular proteins are damaged. Trypsinization must therefore be stopped.

The specially selected proteolytic and collagenolytic enzymes of our Accutase/Biotase solution specifically cleave peptide bonds of extracellular adhesion proteins and show a much gentler cell separation profile than trypsin. Inactivation of the enzymes is therefore not necessary, simply dilute the cell suspension to stop detachment.

Comparison of Accutase vs. Trypsin

• Gentle detachment: Accutase detaches cells more gently, causing less cell damage and less disruption to the cell membrane compared to trypsin.
• Higher viability: Thanks to the gentler process, a larger proportion of cells remain viable and metabolically active.
• Better preservation of surface markers: Accutase-based cell detachment preserves surface proteins, which facilitates subsequent immunophenotyping (e.g., FACS) and functional analyses. Downstream applications such as gene or protein studies are also simplified.
• Broader compatibility with cell types: Accutase is particularly useful for adherent cells that are sensitive to proteolytic activity. Many stem and epithelial cells show improved detachment with Accutase.
• Less activation of stress-induced signaling pathways: Accutase-based cell detachment causes less cellular stress and minimizes the activation of stress signaling pathways.
• Simpler handling: Accutase requires shorter digestion times than trypsin and does not require inactivation. Accutase is therefore practical and flexible.