HK Mad2-LAP/H2B-mCherry

The HK Mad2-LAP/H2B-mCherry cell line is a genetically engineered cell model extensively utilized for studying chromosome segregation and the spindle assembly checkpoint during mitosis. These cells are derived from HeLa Kyoto cells, a robust human cell line originally taken from a cervical carcinoma. The HK Mad2-LAP (LAP-tagged Mad2) aspect of the cell line facilitates the visualization and functional analysis of the Mad2 protein, a critical component of the spindle assembly checkpoint that prevents anaphase onset until all chromosomes are properly aligned at the metaphase plate.

Incorporation of H2B-mCherry, where histone H2B is tagged with the mCherry fluorescent protein, allows for real-time imaging of chromatin dynamics during cell division. This feature makes the HK Mad2-LAP/H2B-mCherry cell line an excellent tool for high-resolution live-cell imaging techniques to observe chromosomal movements and mitotic progression in human cells under various experimental conditions. The use of fluorescent tags aids in precise tracking and quantification, thereby providing valuable insights into the molecular mechanisms governing cell cycle regulation and chromosomal stability.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.