T4424

Sigma-Aldrich

Trypsin solution from porcine pancreas

1 ×, sterile-filtered, BioReagent, suitable for cell culture, 2.5 g porcine trypsin per liter in Hanks′ Balanced Salt Solution with phenol red

CAS Number:
Enzyme Commission number:
MDL number:
NACRES:
NA.75

biological source

Porcine pancreas

Quality Level

sterility

sterile-filtered

product line

BioReagent

form

solution

mol wt

23.4 kDa

concentration

1 ×

application(s)

cell culture | mammalian: suitable

impurities

Porcine parvovirus, none detected (9 CFR)

pH

7.0-7.6

shipped in

dry ice

storage temp.

−20°C

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Application

Trypsin solution from porcine pancreas has been used for keratinocyte isolation and staining. It has also been used as a component of CMF-Steinberg′s solution to replace heart tissues after centrifugation in a study to culture embryonic cardiomyocytes from Mexican axolotl.

For trypsin digestion of peptides, use a ratio of about 1:100 to 1:20 for trypsin:peptide. 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. Trypsins have also been used for the re-suspension of cells during cell culture, in proteomics research for digestion of proteins and in various in-gel digestions. Additional applications include assessing crystallization by membrane-based techniques and in a study to determine that protein folding rates and yields can be limited by the presence of kinetic traps.

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.

Caution

This product should be stored frozen at -20°C

Unit Definition

One BAEE unit will produce a A253 of 0.001 per minute at pH 7.6 at 25°C using BAEE as a substrate.

Preparation Note

This is a 2.5 g/L porcine trypsin solution in Hank′s Balanced Salt Solution with phenol red.

RIDADR

NONH for all modes of transport

WGK Germany

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Certificate of Origin

A method for the isolation and culture of embryonic cardiomyocytes from Mexican axolotl.
R W Zajdel et al.
The International journal of developmental biology, 40(4), 907-908 (1996-08-01)
Ylva Rosengardten et al.
Aging cell, 10(6), 1011-1020 (2011-09-10)
Hutchinson-Gilford progeria syndrome (HGPS or progeria) is a very rare genetic disorder with clinical features suggestive of premature aging. Here, we show that induced expression of the most common HGPS mutation (LMNA c.1824C>T, p.G608G) results in a decreased epidermal population...
Simon Joost et al.
Cell reports, 25(3), 585-597 (2018-10-18)
Epithelial tissues, such as the skin, rely on cellular plasticity of stem cells (SCs) from different niches to restore tissue function after injury. How these molecularly and functionally diverse SC populations respond to injury remains elusive. Here, we genetically labeled...
Colleen Nofi et al.
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 66(2), 99-120 (2017-11-09)
This study was aimed to understand the mechanism of persistent cardiac myocyte (CM) survival in myocardial infarction (MI) scars. A transmural MI was induced in 12-month-old Sprague-Dawley rats by permanent coronary artery ligation. The hearts were collected 3 days, 1...
Inma M Berenjeno et al.
Nature communications, 8(1), 1773-1773 (2017-11-25)
Mutations in PIK3CA are very frequent in cancer and lead to sustained PI3K pathway activation. The impact of acute expression of mutant PIK3CA during early stages of malignancy is unknown. Using a mouse model to activate the Pik3ca
Protocols
Trypsin may be used to remove adherent cells from a culture surface. Cells are most commonly removed from the culture substrate by treatment with trypsin or trypsin/EDTA solutions.
Read More
Trypsin is frequently used in cell dissociation from adherent surfaces. We offer a wide variety of trypsin solutions to meet your specific cell line requirements, as well as protocols, troubleshooting ideas, and more.
Read More

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