Description

PurMa™ Trypsin EDTA Free 0.25%

Background

PurMa™ Trypsin EDTA Free 0.25% contains ultra-pure trypsin. This enzyme is a member of the serine protease family. Trypsin, initially produced in the pancreas, is subsequently secreted and activated in the intestines. Cell culture laboratories use trypsin to dissociate adherent cells for routine cell culture.

Trypsin Activity

Trypsin cleaves its substrates: IGDKLIPA unless these amino acids are followed by proline. PurMa™ trypsin is HPLC purified at 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.

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: 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…

PurMa™ Trypsin Deactivates Mycoplasma in your Cell Culture

Trypsin treatment abolishes the ability of mycoplasmas to attach to the cells. Incidentally, mycoplasma is a common, 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.

Features and Benefits

PurMa Biologics manufactures trypsin without EDTA 0.05 % as a complete Ready-To-Use solution that contains 1X Hanks’ buffer and DOES NOT need any additional ingredients.

• PurMa™ Trypsin EDTA Free 0.25% does not contain calcium or magnesium ions or any other heavy metals.
• PurMa™ Trypsin EDTA Free 0.25% does not contain serine protease inhibitors, including DFP, TLCK, APMSF, and AEBSEF.
• PurMa™ Trypsin EDTA Free 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 is rigorously evaluated for yeast, mold, mycoplasma, and endotoxin contamination.

Concentrations

PurMa Biologics produces 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, we manufacture PurMa™ Trypsin in Hanks’ Buffer with 2mM EDTA (pH 7.8).

Here is the complete formulation: 0.25% Trypsin without EDTA 

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.