人PRKACB (PKA beta catalytic subunit) knockout HeLa cell line

处理步骤

初始处理指南

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO₂. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x10⁴ cells/cm². Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

传代培养指南

• All seeding densities should be based on cell counts gained by established methods.
• A guide seeding density of 2x10⁴ cells/cm² is recommended.
• Cells should be passaged when they have achieved 80-90% confluence.

培养基

DMEM (High Glucose) + 10% FBS

低温储藏试剂

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

质量控制

STR 分析

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

细胞培养

生物安全等级

EU: 2 US: 2

贴壁/悬浮

Adherent

性别

Female