KC-2112

HEK293-AR-Cell-Line

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Background of HEK293-AR-Cell-Line

AR, also known as AIS, belongs to the nuclear receptor family, and modulates a range of processes during embryogenesis and in the adult. During embryogenesis, normal AR function is critical to the development of the male phenotype and defects of the AR cause a range of phenotypic abnormalities of male sexual development. AR is mainly found on prostate cancer, and the emergence of considerable heterogeneity of AR expression at early stages in the development of prostate cancer.

Specifications

Catalog Number	KC-2112
Cell Line Name	HEK293-AR-Cell-Line
NCBI/UniProt Accession Number	NM_000044.6
Clone Number	4#
Host Cell Line	Human HEK293T cell line
Description	Stable HEK293 cell line expressing exogenous human AR 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% DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10%FBS+1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:3-1:6 every 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

HEK293 human AR cell line was generated using a lentiviral vector expressing the human AR sequence.

Characterization

Figure: Characterization of AR overexpression in HEK293 stable clones using Western blot.

Cell Resuscitation

Prewarm culture medium (DMEM + 10% FBS + 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 1:3-1:6 every 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

Application of AR and VR in hand rehabilitation: A systematic review.2020 Nov;111:103584. doi: 10.1016/j.jbi.2020.103584.
The androgen receptor (AR) in syndromes of androgen insensitivity and in prostate cancer.D M Avila 1, S Zoppi, M J McPhaul.Jan-Mar 2001;76(1-5):135-42.
The androgen receptor gene and its influence on the development and progression of prostate cancer.J S Montgomery 1, D K Price, W D Figg.2001 Sep;195(2):138-46.