KC-5967

293T-chimeric-STEAP1-loop1-Flag Cell Line

60639

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Background of 293T-chimeric-STEAP1-loop1-Flag Cell Line

This gene is predominantly expressed in prostate tissue, and is found to be upregulated in multiple cancer cell lines. The gene product is predicted to be a six-transmembrane protein, and was shown to be a cell surface antigen significantly expressed at cell-cell junctions.

Specifications

Catalog Number	KC-5967
Cell Line Name	293T-chimeric-STEAP1-loop1-Flag Cell Line
NCBI/UniProt Accession Number	NM_012449.3
Clone Number	12#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous chimeric-STEAP1-loop1-Flag gene.
Quantity	One vial 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% DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10% FBS +1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 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 30 hours
Mycoplasma Status	Negative

Cell Line Generation

293T-chimeric-STEAP1-loop1-Flag cell line was generated using lentivirus expressing chimeric-STEAP1-loop1-Flag sequence.

Characterization

Figure 1. Characterization of chimeric-STEAP1-loop1-Flag over-expression in the 293T-chimeric-STEAP1-loop1-Flag stable clone using FACS.

Figure 2.Characterization of 293T-chimeric-STEAP1-loop1-Flag cell line stable clone using PCR sequencing.

Cell Resuscitation

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

Cell Freezing

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

References

Zhang Z, Hou WB, Zhang C, Tan YE, Zhang DD, An W, Pan SW, Wu WD, Chen QC, Xu HM. A research of STEAP1 regulated gastric cancer cell proliferation, migration and invasion in vitro and in vivos. J Cell Mol Med. 2020 Dec;24(24):14217-14230. doi: 10.1111/jcmm.16038. Epub 2020 Oct 30. Erratum in: J Cell Mol Med. 2022 Jan;26(1):242-243. doi: 10.1111/jcmm.17117. PMID: 33128353; PMCID: PMC7754049.

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.