KC-2767

293T-cyno-LY6G6D Cell Line

33940

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Background of 293T-cyno-LY6G6D Cell Line

LY6G6D,also known as LY6-D, is a type I transmembrane protein belonging to the immunoglobin (Ig) superfamily, that is comprised of cell-surface proteins involved in the immune system and cellular recognition. And it is known to interact with GRB2 and GRB7 in a phosphorylation-dependent manner thus playing an important role in their downstream signal transduction pathways. Diseases associated with LY6G6D include Glucosephosphate Dehydrogenase Deficiency and Anemia, Nonspherocytic Hemolytic, Due To G6pd Deficiency.

Specifications

Catalog Number	KC-2767
Cell Line Name	293T-cyno-LY6G6D Cell Line
Clone Number	NA
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous cyno-LY6G6D 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 1:4-1:8 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

293T-cyno-LY6G6D cell line was generated using lentivirus expressing cyno-LY6G6D sequence.

Characterization

Figure 1. Characterization of cyno-LY6G6D over-expression in the 293T-cyno-LY6G6D stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 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 1:4-1:8 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

Mallya M, Campbell RD, Aguado B (July 2002). "Transcriptional analysis of a novel cluster of LY-6 family members in the human and mouse major histocompatibility complex: five genes with many splice forms". Genomics. 80 (1): 113–23. doi:10.1006/geno.2002.6794. PMID 12079290.
"Entrez Gene: Lymphocyte antigen 6 complex, locus G6E pseudogene)".
"Human PubMed Reference:". National Center for Biotechnology Information, U.S. National Library of Medicine.