Gibco™ TrypLE™ Express Enzyme (1X), phenol red

Product Code.	Quantity	unitSize
11568856	20 x 100mL	Pack of 20
10043382	100 mL	100mL
11558856	500 mL	500mL

Product Code. 11568856 Supplier Gibco™ Supplier No. 12605036

Description

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. Exceptional purity increases specificity and reduces damage to cells that can be caused by other enzymes present in some trypsin extracts.

• Gentle On Cells
• urity increases specificity due to the action of single enzyme, reducing damage caused by cleavage from enzymes present in some trypsin preparations and in other protease extracts
• Room-Temperature Stable for 24 months
• Ready to use
• Can be directly substituted for trypsin in existing protocols
• Dilution alone inactivates TrypLE Express, avoiding the need for trypsin inhibitors, such as FBS

Energy efficient, less hazardous, sustainable packaging

Cell Culture, Mammalian Cell Culture

Order Info

Shipping Condition: Room Temperature

Compliance

Manufactured in facilities compliant with cGMP manufacturing requirements, certified to the ISO 13485 standard, and registered with the FDA as medical device manufacturers.

Specifications

• 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
• With Additives: Phenol Red, EDTA
• Osmolality: Check and Record
• Product Line: Gibco
• Quantity: 20 x 100mL
• Remarks: Room Temperature Stable
• Shipping Condition: Room Temperature
• Source: Animal origin-free
• Tested For: In vitro Bioassay

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 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.

Safety and Handling

Product Identifier

• 1-(3-Methoxyphenyl)piperazine dihydrochloride

Signal Word

• Warning

Hazard Category

• Serious eye damage/eye irritation Category 2
• Skin corrosion/irritation Category 2
• Specific target organ toxicity Category 3

Hazard Statement

• H315-Causes skin irritation.
• H319-Causes serious eye irritation.
• H335-May cause respiratory irritation.

Precautionary Statement

• P280-Wear protective gloves/protective clothing/eye protection/face protection.
• P302+P352-IF ON SKIN: Wash with plenty of water/soap.
• P304+P340-IF INHALED: Remove person to fresh air and keep comfortable for breathing.
• P312-Call a POISON CENTER/doctor/if you feel unwell.
• P332+P313-If skin irritation occurs: Get medical advice/attention.
• P337+P313-If eye irritation persists: Get medical advice/attention.

Supplemental information

• MIXTURE LIST-Contain: 1-(3-Methoxyphenyl)piperazine dihydrochloride

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