Worthington Dissociation Enzymes: Trypsin and Frequently Asked Questions

A basic overview of Ai Meiji Worthington:

Trypsin is a pancreatic serine protease with specificity for peptide bonds involving the carboxyl groups of basic amino acids, arginine and lysine.Trypsin is one of the most specific proteases known, although it also exhibits some esterase and amidase activities.

Purified trypsin alone is generally ineffective for tissue dissociation because it has little selectivity for extracellular proteins.Combinations of purified trypsin and other enzymes such as elastase and/or collagenase have proven effective for dissociation.

"Trypsin" is also the commercial supplier's name for trypsin, a crude mixture of proteases, polysaccharases, nucleases, and lipases extracted from porcine pancreas.NF 1:250 is a commonly used preparation of "trypsin" that has the ability to proteolytically digest 250 times its weight of casein under assay conditions specified in the National Formulary.It is important to realize that this assay procedure is not specific for trypsin, although trypsin does contain this enzyme.Despite the nomenclature, crude "trypsin" like NF 1:250 and 1:300 are widely used to dissociate tissue, probably because trypsin and contaminating proteolytic and polysaccharidase activities do attack preferentiallyextracellular matrix.

In a tissue culture laboratory, researchers use purified trypsin to release cells into suspension from a monolayer grown on the inner surface of a culture vessel.Most cells derived from normal tissues and not highly adapted to artificial culture conditions grow as a monolayer, a layer of cells one cell thick that adheres to the inner surface of the culture vessel.Because such cells are more like cells in normal tissues, many tissue culture researchers are studying cells grown in monolayer cultures.

Monolayer cultures are typically grown in glass or polystyrene roller bottles, culture flasks, or petri dishes.Plastic containers used for tissue culture work are specially treated to ensure that cells adhere well to the container walls.

Some of the J commonly used purified trypsin grades for cell isolation procedures are EmtechyWorthingtoncodes: TL, TRL, TRLS, and TRLVMF.These are suitable for cell harvesting as well as tissue dissociation.

Organization table (reference, grouped by tissue type and species)

Fat/fat adrenal bone brain

Cartilage Colon Endothelial Epithelium

Eyes Heart Intestines Kidneys

Liver, Lung, Lymph node, Breast

Various muscles, nerves, pancreas

Parotid Pituitary Prostate Reproduction

Scale skin dry spleen

thymus thyroid/parathyroid tonsil tumor

Note: In general, tissue dissociating enzymes from Worthington are used interchangeably for most of the formulations cited.

Worthington core enzymes: including: collagenase, deoxyribonuclease I, elastase, papain, ribonuclease, trypsin, etc.