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TrypLE™ Select (10X) - FAQs

View additional product information for TrypLE™ Select (10X) - FAQs (0040090DG)

5 product FAQs found

Is there an upper limit for the amount of time cells can be incubated with TrypLE Express Enzyme, TrypLE Select Enzyme, or CTS TrypLE Select Enzyme for dissociation?

There is no upper limit for TrypLE enzyme incubation. You can incubate your cells with TrypLE longer to detach cells. 30-45 min at 37 degrees C will be usually fine. Or you can consider using TrypLE Select Enzyme (10X) (Cat. No. A1217701 or A1217702 or A1217703) or CTS TrypLE Select Enzyme (Cat. No. A4738001) depending upon the application.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

For your TrypLE products, how does the enzyme get inactivated?

Our TrypLE enzymes are very gentle, and there is no need to use serum or individual proteins to inactivate the enzyme. The TrypLE enzyme activity is inactivated by dilution using the following procedure:

After cell dissociation, transfer the cell suspension to a 15-mL conical tube. Centrifuge at 100 x g for 5-10 min to remove residual enzyme activity. Discard the supernatant and resuspend the cell pellet in 2-5 mL of pre-warmed complete medium.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Why are my cells not adhering when I transfer them to a new flask?

Trypsinize this type of cells for a shorter duration or use less trypsin or Gibco TrypLE Select or Gibco TrypLE Express Trypsin Replacement. After trypsinization dilute in 2 to 5 mL of cell culture growth medium transfer the cell suspension to 15 mL centrifuge tube. Centrifuge for 5-10 minutes at 100 x g. Discard the supernatant and suspend cell pellet in 2-5 mL fresh medium. Determine the count and seed the flask according to normal protocol and cell should adhere to the surface.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Do you have a non-animal-origin enzyme for cell dissociation?

Yes, the TrypLE products contain rProtease, a non-animal, trypsin-like enzyme used for the dissociation of attachment dependent cell lines. TrypLE enzyme has demonstrated the ability to dissociate cells cultured both in serum-free and serum-supplemented systems. The catalog numbers for TrypLE Select are 12563-011 (100 mL) and 12563-029 (500 mL). Some example catalog numbers for TrypLE Express include 12604-013 without Phenol red (100 mL) and 12605-010 with Phenol red (100 mL). Both formulations are also available in additional sizes. All these products are animal origin-free.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

What can be used to detach cells from plates when coated with a thin layer of collagen I?

Enzymes that can be used include: Gibco TrypLE Express Enzyme, Gibco TrypLE Select Enzyme, or Gibco StemPro Accutase Cell Dissociation Reagent.

Find additional tips, troubleshooting help, and resources within our Extracellular Matrices (ECM) Support Center.