KC-4295

CHOK1-STEAP1B2 Cell Line

44226

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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	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% 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.

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.