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Gibco™ TrypLE™ Express Enzyme (1X), no phenol red
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Artikelnummer. 11538856 Lieferant Gibco™ Lieferanten-Nr. 12604039
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Beschreibung

Unlike porcine trypsin, free of animal-derived components

TrypLE Express is an animal origin-free, recombinant enzyme used for dissociating a wide range of adherent mammalian cells, including CHO, HEK 293, A529, primary human keratinocytes, and embryonic stem cells. TrypLE Express cleaves peptide bonds on the C-terminal sides of lysine and arginine, and is a direct replacement for trypsin. Its exceptional purity increases specificity and reduces damage to cells that can be caused by other enzymes present in some trypsin extracts. We offer a variety of TrypLE formulations for a range of cell culture applications.

Compared to trypsin and other dissociation reagents, TrypLE Express is:

  • Gentle on cells
  • Room temperature-stable
  • Easy to use
  • Animal origin-free (AOF)

This TrypLE Express is modified as follows:

With: EDTA

Without: Phenol Red

The formulation of TrypLE solutions is available, although we keep the enzyme concentration confidential.

Gentle on cells

TrypLE Express' exceptional purity increases specificity due to the action of a single enzyme. This reduces damage caused by cleavage from multiple enzymes in trypsin and other extracts.

Room temperature-stable

TrypLE Express remains stable for 24 months at room temperature or 2–8°C, making storage and handling easy and convenient.

Easy to use

TrypLE Express can be directly substituted for trypsin in existing protocols. In addition, dilution alone inactivates TrypLE Express, avoiding the need for trypsin inhibitors, such as FBS.

Animal origin-free (AOF)

Unlike porcine trypsin, TrypLE Express is free of animal-derived components.

cGMP manufacturing and quality system

For supply chain continuity, we manufacture Gibco TrypLE Express Enzyme (1X), no phenol red at two separate facilities located in Grand Island, NY and Scotland, UK. Both sites are compliant with cGMP manufacturing requirements and are certified to the ISO 13485 standard.

Bestellinformation

Shipping Condition: Room Temperature

Compliance

Manufactured at a cGMP-compliant facility that is registered with the FDA as a medical device manufacturer and is certified to ISO 13485 standards.

TRUSTED_SUSTAINABILITY

Spezifikation

Chelators EDTA
Content And Storage Storage conditions: 15°C to 30°C, protect from light
Alternate storage conditions: 2°C to 8°C or -5°C to -20°C
Shipping conditions: Room temperature
Shelf life: 24 months from date of manufacture
Cell Type Mammalian Cells
Classification Animal Origin-free
Concentration 1 X
Culture Type Adherent Cell Culture
Form Liquid
Product Type Cell Culture Dissociation Reagent
Sterility Sterile-filtered
Without Additives No Phenol Red
Green Features ACT Ecolabel (Silver)
Osmolality Check and Record
pH 6 to 8
Product Line Gibco
Quantity 20 x 100mL
Remarks Room Temperature Stable
Shipping Condition Room Temperature
Source Animal origin-free
Tested For In vitro Bioassay
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Frequently Asked Questions (FAQs)

What enzymes do you recommend for passaging mesenchymal stem cells (MSC)?

We recommend using TrypLE Express Enzyme or TrypLE Select Enzyme for passaging mesenchymal stem cells (MSCs). The usage of trypsin will result in lower cell viability upon passage. TrypLE Express and TrypLE Select are defined, animal origin-free, and stable at both room and refrigerated temperatures.

Does Diploid Growth Serum-Reduced Medium (SRM) contain L-glutamine?

The basal Diploid Growth Serum-Reduced Medium already contains 6 mM L-glutamine.

Do I have to adjust my infection parameters when using Diploid Production Serum-Free Medium (SFM)?

We recommend evaluating performance with Diploid Production Serum-Free Medium using current conditions, however, multiplicity of infection, time of infection, and time of harvest may be different than with conventional media. Closely monitor cells for cytopathic effect and evaluate viral titers at multiple time points.
Can Diploid Growth Serum-Reduced Medium (SRM) be supplemented with different types of serum?

Diploid Growth Serum-Reduced Medium can be supplemented with Fetal Bovine Serum, Newborn Calf Serum, Donor Bovine Serum with Iron, or Bovine Serum.
Which cell lines can I culture in Diploid Growth Serum-Reduced Medium (SRM)?

Diploid Growth Serum-Reduced Medium has been evaluated with fibroblast cell lines, including MRC-5, WI-38, IMR-90, BS-2, and CEF cells.
How long can I use Diploid Growth Serum-Reduced Medium (SRM) supplemented with Diploid Growth Supplement?

Once supplemented, the complete Diploid Growth Serum-Reduced Medium must be used within 4 weeks.
Can I freeze or thaw my cells in Diploid Growth Serum-Reduced Medium (SRM)?

Most diploid cell lines can be frozen and thawed in Diploid Growth Serum-Reduced Medium. Consult the Diploid Growth Serum Reduced Medium User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0019015_DiploidGrowthSerumReducedMedium_UG.pdf) for additional information.
How do I adapt my cells to Diploid Growth Serum-Reduced Medium (SRM)?

Cells growing in medium with serum can be transferred directly to Diploid Growth Serum-Reduced Medium without adaptation. Continue to monitor and passage cells for 2-3 passages until consistent growth is achieved. Cultures may benefit from supplementation with 1-2% serum.
Does TrypLE Express Enzyme have a His-tag?

TrypLE Express Enzyme does not have a His-tag
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.

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.

What is the difference between TrypLE Express Enzyme, with and without phenol red? What does the phenol red do?

The difference between the two products is just the presence or absence of phenol red. Phenol red is a pH indicator. You can use either product to dissociate your cells. When you use TrypLE Express enzyme to dissociate cells, they very likely become single cells. If you want to passage cells as clumps, you may consider using other reagents such as EDTA.

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.

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.
What is the difference between TrypLE Express Enzyme, TrypLE Select Enzyme, and CTS TrypLE Select Enzyme?

All the above products contain rProtease, a non-animal, trypsin-like enzyme formulation used for the dissociation of attachment-dependent cell lines. This alternative for porcine trypsin is a recombinant enzyme derived from microbial fermentation.

The TrpLE Express, TrypLE Select, and CTS TrypLE Select products differ in their production and testing methods and in the documentation provided.

TrypLE Select is for the industrial market and has release-testing both for endotoxins and enzyme activity. This material also has a Drug Master File on record. It is manufactured in AOF-dedicated equipment and is intended for Research Use/Further Cell Culture Manufacturing.

TrypLE Express is manufactured without use of dedicated equipment and is intended for Research Use Only. Not for use in diagnostic procedures.

CTS TrypLE Select is manufactured under scalable cGMP conditions using dedicated equipment and is designed for clinical applications. This product has a Drug Master File on record and is intended for Research Use or Manufacturing of Cell, Gene, or Tissue-Based Products. Not intended for direct administration into humans or animals.

To read more about TrypLE and cell release times using this reagent, please go here (https://www.thermofisher.com/us/en/home/life-science/cell-culture/mammalian-cell-culture/reagents/trypsin/tryple-express.html#1).

What are the benefits of Gibco TrypLE reagent over trypsin?

TrypLE reagent is free of animal- and human-derived components, producing an exceptionally pure reagent that is gentle on cells. Inactivation with trypsin inhibitors is not required. TrypLE reagent is ideal for dissociating a variety of attachment-dependent mammalian cell lines both in serum and in serum-free conditions, and can be directly substituted for trypsin in your current protocol. TrypLE reagent is room-temperature stable and ready to use when you need it. TrypLE cell dissociation reagents remain stable for 24 months at room temperature, making storage and handling easier and more convenient.
What reagents do you offer for cell dissociation, and what are the main differences between them?

Please use this selection chart that compares our cell dissociation reagents (https://www.thermofisher.com/us/en/home/life-science/cell-culture/mammalian-cell-culture/reagents/trypsin.html).

Do you offer media without phenol red (phenol red-free media)?

Yes, please visit this page for a list of our phenol red-free media (http://tools.thermofisher.com/content/sfs/brochures/Phenol_RedFree_Media_Glance.pdf).
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.

For Research Use Only. Not for use in diagnostic procedures.

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