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T4299 Sigma

Trypsin-EDTA solution

1 ×, sterile-filtered, BioReagent, suitable for cell culture, 500 BAEE units porcine trypsin and 180 μg EDTA • 4Na per ml in Dulbecco′s PBS without calcium and magnesium




Trypsin consists of a single chain polypeptide of 223 amino acid residues, produced by the removal of the N-terminal hexapeptide from trypsinogen which is cleaved at the Lys - lle peptide bond. The sequence of amino acids is cross-linked by 6 disulfide bridges. This is the native form of trypsin, beta-trypsin. BETA-trypsin can be autolyzed, cleaving at the Lys - Ser residue, to produce alpha-trypsin. Trypsin is a member of the serine protease family.

Biochem/physiol Actions

Trypsin cleaves peptides on the C-terminal side of lysine and arginine residues. The rate of hydrolysis of this reaction is slowed if an acidic residue is on either side of the cleavage site and hydrolysis is stopped if a proline residue is on the carboxyl side of the cleavage site. The optimal pH for trypsin activity is 7-9. Trypsin can also act to cleave ester and amide linkages of synthetic derivatives of amino acids. EDTA is added to trypsin solutions as a chelating agent that neutralizes calcium and magnesium ions that obscure the peptide bonds on which trypsin acts. Removing these ions increases the enzymatic activity.

Serine protease inhibitors, including DFP, TLCK, APMSF, AEBSEF, and aprotinin, amongst others, will inhibit Trypsin.


This product is stored frozen between -10 and -40°C. Repeated cycles of freezing and thawing should be avoided.


The typical use for this product is in removing adherent cells from a culture surface. The concentration of trypsin necessary to dislodge cells from their substrate is dependent primarily on the cell type and the age of the culture.

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Safety & Documentation

Safety Information

WGK Germany 


Certificate of Analysis

Protocols & Articles
Peer-Reviewed Papers


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Pivotal roles of lymphatic endothelial cell layers in the permeability to hydrophilic substances through collecting lymph vessel walls: effects of inflammatory cytokines. Kawai Y, Kaidoh M, Yokoyama Y, et al. Lymphat. Res. Biol. 12(3), 124-35, (2014)


Role of Drosophila retinoblastoma protein instability element in cell growth and proliferation. Elenbaas JS, Mouawad R, Henry RW, et al. Cell Cycle 14(4), 589-97, (2015)


Coherent but overlapping expression of microRNAs and their targets during vertebrate development. Shkumatava A Genes Dev. 23(4), 466-81, (2009)


The generation of live offspring from vitrified oocytes. L. Gabriel Sanchez-Partida et al PLoS ONE 6, e21597, (2011)


A new potent secondary amphipathic cell-penetrating peptide for siRNA delivery into mammalian cells. Laurence Crombez et al Mol. Ther. 17, 95-103, (2008)


Effect of processed and red meat on endogenous nitrosation and DNA damage. Joosen AM Carcinogenesis 30(8), 1402-7, (2009)


TrkB regulates hippocampal neurogenesis and governs sensitivity to antidepressive treatment. Li Y, Luikart BW, Birnbaum S, et al. Neuron 59(3), 399-412, (2008)


Modulation of human embryonic stem cell-derived cardiomyocyte growth: a testbed for studying human cardiac hypertrophy? Gábor Földes et al J. Mol. Cell. Cardiol. 50, 367-76, (2011)


New evidence confirms that the mitochondrial bottleneck is generated without reduction of mitochondrial DNA content in early primordial germ cells of mice. Cao L PLoS Genet. 5(12), e1000756, (2009)


Imidazole metalloporphyrins as photosensitizers for photodynamic therapy: role of molecular charge, central metal and hydroxyl radical production. Mroz P Cancer Lett. 282(1), 63-76, (2009)


Contribution of human muscle-derived cells to skeletal muscle regeneration in dystrophic host mice. Jinhong Meng et al PLoS ONE 6, e17454, (2011)


3,3'-Diindolylmethane stimulates murine immune function in vitro and in vivo. Ling Xue et al J. Nutr. Biochem. 19, 336-44, (2008)


TOPAZ1, a novel germ cell-specific expressed gene conserved during evolution across vertebrates. Baillet A, Le Bouffant R, Volff JN, et al. PLoS ONE 6(11), e26950, (2011)


Enhanced growth and osteogenic differentiation of human osteoblast-like cells on boron-doped nanocrystalline diamond thin films. Grausova L PLoS ONE 6(6), e20943, (2011)


ncreased expression of 5-HT(1B) receptors by Herpes simplex virus gene transfer in septal neurons: New in vitro and in vivo models to study 5-HT(1B) receptor function. Riegert C., et al. Brain Res. Bull. 76, 439-453, (2008)


FRS2α is essential for the fibroblast growth factor to regulate the mTOR pathway and autophagy in mouse embryonic fibroblasts. Lin, X Int. J. Biol. Sci. 7(8), 1114-21, (2011)


β1-6 branching of cell surface glycoproteins may contribute to uveal melanoma progression by up-regulating cell motility Lityska, A., et al. Mol. Vis. 14, 625-636, (2008)


Spatial separation and bidirectional trafficking of proteins using a multi-functional reporter. Svendsen S BMC Cell Biol. 9, 17, (2008)


The bHLH factors extramacrochaetae and daughterless control cell cycle in Drosophila imaginal discs through the transcriptional regulation of the Cdc25 phosphatase string. Andrade-Zapata I and Baonza A PLoS Genet. 10(3), e1004233, (2014)


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