KC-4295

CHOK1-STEAP1B2 Cell Line

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Background of CHOK1-STEAP1B2 Cell Line

Predicted to be located in membrane. Predicted to be active in endosome and plasma membrane. STEAP1B (STEAP Family Member 1B) is a Protein Coding gene. Gene Ontology (GO) annotations related to this gene include cupric reductase (NADH) activity and ferric-chelate reductase (NADPH) activity. An important paralog of this gene is STEAP1.

Specifications

Catalog Number	KC-4295
Cell Line Name	CHOK1-STEAP1B2 Cell Line
Clone Number	5#
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous STEAP1B2 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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 10μg/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
In Vivo Validation	NA

Cell Line Generation

CHOK1 STEAP1B2 cell line was generated using a lentiviral vector expressing the STEAP1B2 sequence.

Characterization

Figure 1: Characterization of STEAP1B2 overexpression in the CHOK1 STEAP1B2 stable clone using qPCR.

Figure 2: Characterization of STEAP1B2 and its mutants overexpressing in CHOK1 stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10μg/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% RPMI1640 + 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

Gomes IM, Santos CR, Maia CJ. Expression of STEAP1 and STEAP1B in prostate cell lines, and the putative regulation of STEAP1 by post-transcriptional and post-translational mechanisms. Genes Cancer. 2014 Mar;5(3-4):142-51. doi: 10.18632/genesandcancer.13. PMID: 25053991; PMCID: PMC4091532.
Murea M, Lu L, Ma L, Hicks PJ, Divers J, McDonough CW, Langefeld CD, Bowden DW, Freedman BI. Genome-wide association scan for survival on dialysis in African-Americans with type 2 diabetes. Am J Nephrol. 2011;33(6):502-9. doi: 10.1159/000327985. Epub 2011 May 5. PMID: 21546767; PMCID: PMC3202959.