KC-6299

293T-ADGRL2-Low Cell Line

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Background of 293T-ADGRL2-Low Cell Line

ADGRL2 (Adhesion G Protein-Coupled Receptor L2) is a Protein Coding gene. The encoded protein participates in the regulation of exocytosis. The proprotein is thought to be further cleaved within a cysteine-rich G-protein-coupled receptor proteolysis site into two chains that are non-covalently bound at the cell membrane. Diseases associated with ADGRL2 include Encephalitozoonosis and Eosinophilic Meningitis.

Specifications

Catalog Number	KC-6299
Cell Line Name	293T-ADGRL2-Low Cell Line
NCBI/UniProt Accession Number	NM_001297704.1
Clone Number	12#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous human ADGRL2 gene in low level
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% EMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10% FBS +1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
Subculture	Split saturated culture 1:4-1:8 every 2-3 days
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative

Cell Line Generation

293T-ADGRL2-Low cell line was generated using a lentiviral vector expressing the human ADGRL2 sequence.

Characterization

Figure 1: Characterization of human ADGRL2 overexpression in the 293T-ADGRL2-Low stable clone using FACS.

Figure 2: Characterization of human ADGRL2 in the 293T-ADGRL2-Low stable clone using PCR sequencing.

Cell Resuscitation

Pre-warm complete culture medium (DMEM+10% FBS +1μg/mL Puromycin) in a 37°C water bath.
Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
Incubate the flask in a 37°C in a humidified 5% CO₂ incubator.
Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
Subculture the cells at a ratio of 1:4-1:8 every 2-3 days upon reaching 80%–90% confluency.

Cell Freezing

Prepare the freezing medium (70% basal medium, 20% FBS and 10% DMSO) freshly before use.
Pre-chill the freezing medium on ice and label the cryovials accordingly.
Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
Gently resuspend the cell pellet in chilled freezing medium, ensuring a minimum cell density of 3×10⁶ cells/mL.
Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

References

1. Jakobs P, Rafflenbeul A, Post WB, Ale-Agha N, Groß VE, Pick S, Dolata S, Cox FF, von Ameln F, Eckermann O, Altschmied J, Prömel S, Haendeler J. The Adhesion GPCR ADGRL2/LPHN2 Can Protect Against Cellular and Organismal Dysfunction. Cells. 2024 Nov 5;13(22):1826. doi: 10.3390/cells13221826. PMID: 39594576; PMCID: PMC11592504.
2. Sarmoko, Toriyama M, Kaji N, Itoh H. Functional Analysis of Adhesion GPCR Latrophilin 2 (ADGRL2) in MDA-MB-231 Human Breast Cancer Cells. Genes Cells. 2025 Jul;30(4):e70030. doi: 10.1111/gtc.70030. PMID: 40545252.
3. Yang X, He F, Lopez-Pajares V, Porter DF, Garbett K, Siprashvili Z, Ducoli L, Meyers RM, Reynolds DL, Lan Huong Bui D, Hong A, Nguyen DT, Jing Y, Mondal S, Ko L, Tao S, Singal B, Sando R, Skiniotis G, Khavari PA. The adhesion GPCR ADGRL2 engages Gα13 to enable epidermal differentiation. Proc Natl Acad Sci U S A. 2025 Nov 25;122(47):e2508436122. doi: 10.1073/pnas.2508436122. Epub 2025 Nov 18. PMID: 41252157; PMCID: PMC12663980.
4. Vezain M, Lecuyer M, Rubio M, Dupé V, Ratié L, David V, Pasquier L, Odent S, Coutant S, Tournier I, Trestard L, Adle-Biassette H, Vivien D, Frébourg T, Gonzalez BJ, Laquerrière A, Saugier-Veber P. A de novo variant in ADGRL2 suggests a novel mechanism underlying the previously undescribed association of extreme microcephaly with severely reduced sulcation and rhombencephalosynapsis. Acta Neuropathol Commun. 2018 Oct 19;6(1):109. doi: 10.1186/s40478-018-0610-5. PMID: 30340542; PMCID: PMC6195752.

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.