KC-6852

293T-mouse-BTN1A1 Cell Line

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Background of 293T-mouse-BTN1A1 Cell Line

BTN1A1 (Butyrophilin Subfamily 1 Member A1) is a Protein Coding gene. Butyrophilin is the major protein associated with fat droplets in the milk. It is a member of the immunoglobulin superfamily. It may have a cell surface receptor function. The human butyrophilin gene is localized in the major histocompatibility complex (MHC) class I region of 6p and may have arisen relatively recently in evolution by the shuffling of exons between 2 ancestral gene families.

Specifications

Catalog Number	KC-6852
Cell Line Name	293T-mouse-BTN1A1 Cell Line
NCBI/UniProt Accession Number	NM_013483.3
Clone Number	4#
Host Cell Line	293T
Description	293T cell line stably expressing exogenous mouse-BTN1A1 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

Cell Line Generation

293T-mouse-BTN1A1 cell line was generated using a lentiviral vector expressing the mouse-BTN1A1 sequence.

Characterization

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 1:4-1:8 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

Heid HW, Winter S, Bruder G, Keenan TW, Jarasch ED. Butyrophilin, an apical plasma membrane-associated glycoprotein characteristic of lactating mammary glands of diverse species. Biochim Biophys Acta. 1983 Feb;728(2):228-38. doi: 10.1016/0005-2736(83)90476-5. PMID: 6830779.
Mather IH, Jack LJ. A review of the molecular and cellular biology of butyrophilin, the major protein of bovine milk fat globule membrane. J Dairy Sci. 1993 Dec;76(12):3832-50. doi: 10.3168/jds.s0022-0302(93)77726-7. PMID: 8132890.
Kim YS, Lee SH, Park AH, Wu C, Hong BK, Jung H, Lin SH, Yoo SS. BTN1A1 is a novel immune checkpoint mutually exclusive to PD-L1. J Immunother Cancer. 2024 Mar 14;12(3):e008303. doi: 10.1136/jitc-2023-008303. PMID: 38485289; PMCID: PMC10941171.

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.