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- 제품/실험 Q&A
Healthy, disease state PBMC 및 Leukopak을 제공합니다.
상세한 characterization, demographic information이 함께 제공됩니다.
|Precision for Medicine|
| Disease State PBMCs
|INSTRUCTIONS FOR THAWING AND CULTURE|
|1. Use a dry ice tub to transfer vials to the lab, making sure to keep the vials submerged in the dry ice to keep them cold.
2. Hold the frozen vial in water in a 37 ± 3°C water bath. Do not thaw more than 4 vials at a time.
3. GENTLY shake the vial in the water until just before the last ice crystal has melted.
4. Check to make sure the cap is secure. It is possible that the cap may loosen slightly with the temperature change between the ice and the water bath.
5. Just before the last ice crystals have melted, remove the vial from the water bath.
6. Wipe the vial with a sterile alcohol pad, focusing on the cap area.
7. Unscrew the cap slowly as pressure may build inside tube on thaw.
8. Add 1 mL of culture media that has been brought to 37 ± 3°C (warm) to the vial drop by drop over 30 seconds to allow the cells to adjust to the change in environment.
9. Slowly add the cells to a 15 or 50 mL conical tube containing 9 mL of warm culture media.
10. Rinse the original vial with 1 mL of the cell-containing media to recover cells that may have adhered to the sides; add the rinse media to the conical tube.
11. Pellet the cells by centrifugation at 350 x g for 8-12 minutes.
12. Discard the supernatant. Re-suspend the cell pellet by gently tapping (avoid excessive shear forces).
13. Rinse the cells again by adding 10 mL of warm culture media to the conical tube.
14. Pellet the cells by centrifugation at 350 x g for 8-12 minutes.
15. Discard the supernatant from the second wash. Re-suspend the cell pellet by gently tapping (avoid excessive shear forces).
16. Resuspend the cells in 5-10 mL of warm media as required.
17. Count cells using laboratory specific procedures and proceed with laboratory protocols.
18. For assays requiring PBMC growth, better results may be obtained if the cells are rested overnight before use.
19. Place the cells in a 50 mL tube, no more than 10 mL per tube.
20. Slightly loosen the cap and place the tubes in a 37°C incubator overnight with the appropriate CO2 concentration.
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