Stem Cell Qualified ECM Gel
Catalog No: HG008
Size: 5 mL
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| Product Description | Below are general guidelines for the coating of 6-well plates using the Stem Cell Qualified ECM Gel. All procedure should be performed under aseptic conditions in a biological safety cabinet. 1. Thaw the Stem Cell Qualified ECM Gel in the 2-8 °C fridge one day before use. Once thawed, maintain ECM Gel on ice at all times and use pre-cooled medium and pipettes to avoid gelling of the product. IMPORTANT: Do not thaw ECM Gel at temperatures higher than 15 °C to avoid gelling. 2. Dilute the ECM Gel 1:80 with cold DMEM/F12 medium. Scale up or down according to the volumes required. Below is an example for coating a 6-well plate with 1.5 mL of 1:80 diluted ECM Gel. • To make a 1:20 dilution, add 0.5 mL ECM Gel to 9.5 mL cold DMEM/F12 or DMEM medium into a 15 mL conical tube. Total volume = 10 mL. • A further 4-fold dilution is required to make the final 1:80 dilution. Add 2.5 mL of 1:20 diluted ECM Gel to 7.5 mL cold DMEM/F12 medium. Total volume = 10 mL NOTE: The recommended dilution is 1:80, however more concentrated Stem Cell Qualified ECM Gel may be used if desired. 3. Add 1.5 mL of the 1:80 diluted Stem Cell Qualified ECM Gel to each well of a 6-well plate. Swirl the culture plates to spread the ECM Gel evenly across the surface of the plate. Store in a 2-8 °C fridge overnight or at least 2 hours in the fridge before use. If not used immediately, parafilm wrap the ECM coated culture plates and store at 2-8 °C until ready to use. Use the ECM coated culture plates within 3-4 days. 4. Prior to seeding the cells, bring the plate back to room temperature for 10-15 minutes, remove the coating solution and add 3 mL/well of human ES/iPSC growth media. Cells can now be plated onto the newly coated plates. IMPORTANT: Do not allow the plates to dry out. |
| Application | Stem cell qualified basement membrane extract (BME) for human ES and iPS cell culture. |
| State | Gel |
| Storage/Stability | Store at -20 °C. Thaw overnight on ice in a 2-8 °C refrigerator. Minimize freeze thaws by aliquoting into one-time use aliquots. Store aliquots in either a -20 °C or -80 °C freezer until ready for use. Do not store in a frost-free freezer. |
| Note | Stem Cell Qualified ECM Gel is extremely temeprature sensitive and may polymerize at room temperature or higher temperature within 5-10 minutes. Keep on ice at all times and use prechilled pipette tips, tubes and plates. |
! For Research/Industry Use Only!