KC-4837

CT26-ERBB2-GFP-Luc2-Cell-Line

56707

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Background of CT26-ERBB2-GFP-Luc2-Cell-Line

ERBB2, also known as CD340, encodes a member of the epidermal growth factor (EGF) receptor tyrosine kinase family. This protein itself does not have a ligand binding domain, so it cannot bind to growth factors. However, it does tightly bind with other ligands in the EGF receptor family to form heterodimers, stabilizing ligand binding and enhancing activation of downstream signaling pathways mediated by kinases, such as those involving mitogen activated protein kinase and phosphatidylinositol 3-kinase. It is reported that the amplification and/or overexpression of this gene are found in many cancers, including breast cancer and ovarian tumors.
Luciferase is an oxidative enzyme that can produce bioluminescence with addition of luciferin, but doesn’t need an external light source unlike fluorescent proteins. Photo emission can be detected directly by light sensitive devices. Such as luminometers or modified microscopes. Luciferase is widely used in many fields of biological research, such as transcriptional activity, kinase or other enzyme activity, cellular ATP level, and whole animal imaging.

Specifications

Catalog Number	KC-4837
Cell Line Name	CT26-ERBB2-GFP-Luc2-Cell-Line
Clone Number	26-8#
Host Cell Line	CT26
Description	Stable CT26 clone expressing exogenous ERBB2 and GFP and Luciferase gene.
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70%RPMI 1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI 1640 + 10% FBS
Selection Marker	NA
Morphology	Fibroblast
Subculture	Split saturated culture 1:4-1:10 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 24 hours
Mycoplasma Status	Negative

Cell Line Generation

CT26-ERBB2-GFP-Luc2-cell-line was generated using a lentiviral vector expressing the ERBB2-GFP-Luc2 sequence.

Characterization

Figure 1: Characterization of ERBB2 overexpression in the CT26-ERBB2-GFP-Luc2 stable clone using FACS.

Figure 2: Characterization of GFP overexpression in the CT26-ERBB2-GFP-Luc2 stable clone using FACS.

Figure 3: Characterization of the CT26-ERBB2-GFP-Luc2-cell-line stable clone using Bright-Lite Luciferase Assay System in the conditions of different cell number.

Figure 4: Characterization of ERBB2 in the CT26-ERBB2-GFP-Luc2 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (RPMI 1640 supplemented with 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 1:4-1:10 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70%RPMI 1640 + 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.

References

1.Greer LF, Szalay AA (2002). Imaging of light emission from the expression of luciferases in living cells and organisms: a review. Luminescence. 17 (1): 43–74. doi:10.1002/bio.676. PMID 11816060.
2.Idamakanti M, Bijjam RI, Kumar M, Mukkamalla SK. Long Durable Response With Trastuzumab Deruxtecan Monotherapy in a Triple-Negative Metastatic Breast Cancer Patient With Human Epidermal Growth Factor Receptor 2 Mutation: A Long-Term Follow-Up and Literature Review. J Med Cases. 2025 Jun 16;16(6):212-221. doi: 10.14740/jmc5136. PMID: 40642740; PMCID: PMC12239829.
3.Zheng Y, Shen G, Zhang C, Huo X, Xin Y, Fang Q, Guan Y, Zhao F, Ren D, Liu Z, Wang M, Zhao J. Efficacy of anti-HER2 drugs in the treatment of patients with HER2-mutated cancers: a systematic review and meta-analysis. Clin Exp Med. 2023 Nov;23(7):3205-3216. doi: 10.1007/s10238-023-01072-7. Epub 2023 Apr 30. PMID: 37120775.