KC-5441

CHOK1-rat-SLC39A6-Cell-Line

56075

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Background of CHOK1-rat-SLC39A6-Cell-Line

SLC39A6 (Solute Carrier Family 39 Member 6) is a Protein Coding gene. Diseases associated with SLC39A6 include Acrodermatitis Enteropathica and Breast Cancer. Among its related pathways are Metal ion SLC transporters and Transport of inorganic cations/anions and amino acids/oligopeptides. Gene Ontology (GO) annotations related to this gene include metal ion transmembrane transporter activity and zinc ion transmembrane transporter activity. An important paralog of this gene is SLC39A10

Specifications

Catalog Number	KC-5441
Cell Line Name	CHOK1-rat-SLC39A6-Cell-Line
Clone Number	12#
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous rat-SLC39A6 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 + 10μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:6-1:8 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 24 hours
Mycoplasma Status	Negative

Cell Line Generation

CHOK1-rat-SLC39A6-cell-line was generated using a lentiviral vector expressing the rat-SLC39A6 sequence.

Characterization

Figure 1: Characterization of rat-SLC39A6 overexpression in the CHOK1-rat-SLC39A6 stable clone using qPCR.

Figure 2: Characterization of rat-SLC39A6 in the CHOK1-rat-SLC39A6 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (RPMI 1640 supplemented with 10% FBS and 10μg/mL puromycin)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 1:6-1:8 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.Manning DL, Daly RJ, Lord PG, Kelly KF, Green CD. Effects of oestrogen on the expression of a 4.4 kb mRNA in the ZR-75-1 human breast cancer cell line. Mol Cell Endocrinol. 1988 Oct;59(3):205-12. doi: 10.1016/0303-7207(88)90105-0. PMID: 2903103.
2.Ma X, Ma Q, Liu J, Tian Y, Wang B, Taylor KM, Wu P, Wang D, Xu G, Meng L, Wang S, Ma D, Zhou J. Identification of LIV1, a putative zinc transporter gene responsible for HDACi-induced apoptosis, using a functional gene screen approach. Mol Cancer Ther. 2009 Nov;8(11):3108-16. doi: 10.1158/1535-7163.MCT-08-0772. Epub 2009 Nov 3. PMID: 19887557.