Trypsin 0.25% - EDTA in HBSS
w/o Calcium w/o Magnesium w/ Phenol Red
CAT N° : IN0008-100
Unit/Size：100ml
Sterility : Sterile filtered
Theoretical pH : 7.3 ± 0.3
Osmolality : 300 mOsm/kg ± 10 %
Colour : orange - red, clear solution
Storage conditions : -20°C
Shelf life : 24 months
Composition :Contains phenol red, 2.5g/L pancreatin (1:250) dissolved in HBSS solution, 0.02% EDTA dissolved in HBSS solution, without calcium and magnesium。
Applications :
Trypsin is a porcine pancreas-derived enzyme that is commonly used for the dissociation and disaggregation of anchorage-dependent mammalian cells and tissues. The concentration of trypsin necessary to dislodge non sensitive cells from their substrate, is 0.25%. EDTA, a chelating agent, enhance the enzymatic activity by removing calcium and magnesium ions. These ions obscure the peptide bonds on which trypsin acts as well as enhancing cell to cell adhesion.
Uses :
The Trypsin 0.25% EDTA in HBSS w/o Calcium w/o Magnesium w/ Phenol Red is a ready to use solution.
1. Frozen products can either be thawed in a 37°C water bath or overnight at to 2 to 8°C.
2. Aspirate the spent medium from the culture vessel and discard.
3. Rinse the monolayer with either a small amount of trypsin solution or a calcium and magnesium-free salt solution (as listed below), aspirate, and discard.
Dulbecco’s Phosphate Buffered Saline (DPBS) catalogIN0011
Hank’s Balanced Salt Solution (HBSS)
4. Add enough trypsin solution, prewarmed in a 37°C water bath, to completely cover the cell monolayer.
5. Incubate the flask at 37°C, or for more sensitive cultures, at room temperature or 2 to 8°C.
6. When the trypsinization process is complete, cells will appear rounded upon microscopic examination and the solution in the flask will appear cloudy. Check the flask often to avoid overexposure which can damage the cells.
7. The trypsin should be neutralized either with serum containing medium or trypsin inhibitor. Gently centrifuge the cell suspension and discard the trypsin-containing supernatant.
8. Resuspend the cell pellet with fresh medium and count or culture as desired.
It can only be used for scientific research. It is forbidden to use it for human, animal or other purposes.
