HEK-Blue™ mMincle Cells
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Murine Mincle / NF-κB / SEAP reporter HEK293 cells
3-7 x 10e6 cells
SEAP Reporter 293 cells expressing the murine Mincle gene
HEK-Blue™ mMincle cells stably express the murine Mincle (mMincle) gene, as well as the genes of the Mincle-NF-kB signaling pathway.
They also stably express an NF‑kB-inducible secreted embryonic alkaline phosphatase (SEAP) reporter gene.
These reporter cell lines are activated by Mincle ligands, such as trehalose-6,6-dibehenate (TDB) and heat-killed M. tuberculosis (HKMT). They do not respond to other CLR ligands such as zymosan, a Dectin-1 and TLR2 ligand.
HEK-Blue™ mMincle and HEK-Blue™ Null1-v cells (parental cell line) were stimulated with various CLR agonists: zymosan (100 µg/ml), HKMT (100 µg/m), TDB (10 µg/ml), and TNF-α (10 ng/ml). TNF-α, an NF-kB activator, was used as a positive control. NF‑kB‑induced SEAP activity was assessed using HEK‑Blue™ Detection and by reading the optical density (OD) at 655 nm.
Growth medium: DMEM (4.5 g/l glucose), 10% (v/v) fetal bovine serum (FBS), 50 U/ml penicillin, 50 µg/ml streptomycin, 100 µg/ml Normocin™, 2 mM L-glutamine.
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- 1 vial containing 3-7 x 106 cells
- 100 μl blasticidin (10 mg/ml).
- 100 μl puromycin (10 mg/ml).
- 2 ml HEK-Blue™ CLR Selection (250x) (a solution containing several selection antibiotics)
- 1 ml Normocin™ (50 mg/ml)
- 1 pouch of HEK-Blue™ Detection (cell culture medium for real-time detection of SEAP)
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HEK-Blue™ mMincle cells have been stimulated by various pathogen recognition receptor (PRR) agonists. As expected, Mincle agonists induced the production of SEAP. The cell surface expression of murine Mincle (mMincle) in this cell line has been validated using fluorescence-activated cell sorting (FACS). The stability of this cell line for 20 passages following thawing has been verified.
Specificity of HEK-Blue™ mMincle cells
As their parental cell line, HEK293 cells, express endogenous levels of TLR3, TLR5 and NOD1, HEK-Blue™ mMincle cells will respond to TLR3, TLR5 and NOD1 agonists, such as poly(I:C), flagellin and C12‑iE‑DAP, respectively. In order to identify Mincle-specific responses, we recommend to use HEK-Blue™ Null1-v cells as a control cell line.