NCI-H1048-CDK2-HiBiT-KI-2A2 Cell Line

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Cyclin-dependent kinases (CDKs) are key players in cell cycle regulation. Cyclin-dependent kinase 2 (CDK2) is best known for its key role during cell cycle progression. This cyclin-dependent kinase (CDK) family member is involved in DNA synthesis, G1/S phase transition, and G2 progression modulation. While the monomeric form of CDK2 is inactive, the kinase becomes active, like many other CDKs, when it forms a functional heterodimeric complex with one of its two regulatory partners – Cyclins A and E. CDK2 modulates a variety of oncogenic signaling pathways because it governs the phosphorylation of a wide range of transcription factors: SMAD3, FOXM1, FOXO1, ID2, UBF, NFY, B-MYB, and MYC. Unsurprisingly, the aberrant activation of CDK2, which occurs in many human cancers, leads to uncontrolled cell proliferation during oncogenesis. Likewise, oncogenic processes of several cancers are also associated with high levels of the two regulatory subunits of CDK2. Altogether, both the activity of CDK2 and its regulatory subunits appear to be important components of oncogenesis.


Catalog Number:
Cell Line Name:
NCI-H1048-CDK2-HiBiT-KI-2A2 Cell Line
Host Cell Line:
Human NCI-H1048 cell line
NCI-H1048 cell line stable clone with endogenous HiBiT-tagged CDK2
One vial of frozen cells (5×106 per vial)
Stable in culture over a minimum of 10 passages
Drug screening and biological assays
Freeze Medium:
70% DMEM+20% FBS+10% DMSO
Propagation Medium:
Selection Marker:
Split saturated culture 1:3-1:8 every 3 days; seed out at about 1-3 × 105 cells/mL
37 °C with 5% CO2
Liquid nitrogen immediately upon receiving
Doubling Time:
Approximately 48 hours
Mycoplasma Status:

Cell Line Generation

NCI-H1048-CDK2-HiBiT-KI cell line was generated using the CRISPR method.


Figure 1: Characterization of NCI-H1048-CDK2-HiBiT-KI cell line stable clone using PCR sequencing.
Figure 2: Characterization of NCI-H1048-CDK2-HiBiT-KI cell line stable clone using RT PCR sequencing.
Figure 3: Characterization of NCI-H1048-CDK2-HiBiT-KI cell line stable clone using western blot.
Figure 4: Characterization of NCI-H1048-CDK2-HiBiT-KI Cell Line stable clone using HiBiT lytic detection.


Cell Resuscitation

1. Prewarm culture medium (DMEM + 10% FBS)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO2 incubator.
8. Split saturated culture 1:3-1:8 every 3 days; seed out at about 1-3 × 105 cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×106 cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage


1. Ding L, Cao J, et al. The Roles of Cyclin-Dependent Kinases in Cell-Cycle Progression and Therapeutic Strategies in Human Breast Cancer. Front Int J Mol Sci (2020 Mar 13): 21(6): 1960.
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