Description

PurMa™ Trypsin-EDTA 0.25%

Background

PurMa™ Trypsin-EDTA 0.25% contains ultra-pure trypsin. Furthermore, trypsin is one of the members of the serine protease family which goes through a set of intracellular signaling results in the activation of this enzyme in pancreas. Cell culture laboratories widely use trypsin to dissociate the adherent cells for routine cell culture. The Trypsin family is conservative. Trypsin performs the best at 37^oC and at the PH 7.3-8.5.

Trypsin Activity

Trypsin cleaves its substrates: IGDKLIPA unless these amino acids are followed by proline. PurMa™ Trypsin is HPLC purified 99% purity for bovine pancreas. Moreover, optical rotatory dispersion of trypsin in the presence of calcium shows a major conformational transition between 40 and 45° which results in  deactivation of trypsin. Therefore, all the buffers needed to be Ca⁺² & Mg⁺²  free for the proper function of this enzyme.

PurMa™ Trypsin Deactivates Mycoplasma in your Cell Culture

Trypsin treatment abolishes the ability of mycoplasmas to attach to the cells. Mycoplasma is one of the common and slowly deteriorating contamination in cell culture, especially due to the invisibility of the contamination. PurMa Biologics manufactures all the reagents you need to fight mycoplasma in your cell culture. The presence of a trace of SDS and other detergents leads to a decrease in the trypsin activity.

Preparation

The activity of this enzyme is further modulated by the sulfation of Tyr154. To overcome the mentioned issue, we use our most sophisticated water quality to ensure the absence of Mg⁺² and Ca⁺² as well as other heavy metals, please see click here: PurMa™ Ultra-Pure Cell Culture Suited Water (Cat# P3W110102)

Additionally, we include 2mM EDTA as it Chelates metals such as calcium, magnesium, lead, mercury, copper, iron…

Features and Benefits

• PurMa™ Trypsin does not contain calcium or magnesium ions or any other heavy metals.
• PurMa™ Trypsin does not contain serine protease inhibitors, including DFP, TLCK, APMSF, AEBSEF.
• PurMa™ Trypsin does not contain Phytates or salts of phytic acid, such as myo-inositol which is commonly present in cell culture media and inhibits trypsin activity.
• PurMa™ Trypsin has been rigorously evaluated for yeast, mold, mycoplasma, and endotoxin contamination.

Concentrations

PurMa Biologics manufactures 0.05%, 0.25%,0.5%, 2.5% and 5% Trypsin with EDTA as well as without EDTA in phenol red or without phenol red. Moreover, PurMa™ Trypsin is made in Hanks’ Buffer with 2mM EDTA (pH 7.8).

Formulation

The complete formulation can be found here: Trypsin with EDTA 0.25%

References

1. Fate of trypsin and chymotrypsin in the human small intestine. Goldberg et al. 1969 Jun;10(6):477-83. doi: 10.1136/gut.10.6.477. PMID: 5786145.
2. Adhesion of mycoplasmas to eukaryotic cells. Razin et al. Ciba Found Symp. 1981;80:98-118. doi: 10.1002/9780470720639.ch8. PMID: 6790254.
3. Effects of hyaluronidase, trypsin, and EDTA on surface composition and topography during detachment of cells in culture. Vogel KG. Exp Cell Res. 1978 May;113(2):345-57. doi: 10.1016/0014-4827(78)90375-0. PMID: 122554.