HEK-Blue™ mMincle Cells

SEAP Reporter 293 cells expressing the murine Mincle gene

HEK-Blue™ mMincle cells have been engineered to express a secreted alkaline phosphatase (SEAP) in response to stimulation by Mincle ligands. 

Mincle is a C-type lectin receptor (CLR) that recognizes a variety of exogenous and endogenous stimuli, such as mycobacteria, certain fungi and necrotic cells [1, 2].

HEK-Blue™ mMincle cells are derived from the human embryonic kidney HEK293 cells which do not naturally express Mincle. We have engineered HEK-Blue™ cells that stably express high levels of murine Mincle as well as the genes of the Mincle-NF-κB signaling pathway. These cells also stably express an NF-κB-inducible SEAP reporter gene. Levels of SEAP are readily measurable in real-time by a simple colorimetric assay with HEK-Blue™ Detection or QUANTI-Blue™ .  

HEK-Blue™ mMincle cells 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.

To identify Mincle-specific responses, we recommend to use HEK-Blue™ Null1-v cells as a control cell line.

HEK-Blue™ hMincle cells expressing the human Mincle gene (hMincle) are also available upon inquiry.

References:

1. Yamasaki S. et al., 2009​. C-type lectin Mincle is an activating receptor for pathogenic fungus, Malassezia. PNAS 106(6): 1897–1902. 
2. Brown GD. 2008.  Sensing necrosis with Mincle. Nature Immunol. 9:1099-1100. 

Figures

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.

Specifications

Required Selective Antibiotics: Blasticidin, Puromycin, and HEK-Blue™ CLR Selection.

Growth medium: DMEM (4.5 g/l glucose), 10% (v/v) fetal bovine serum (FBS), 100 U/ml penicillin, 100 µg/ml streptomycin, 100 µg/ml Normocin™, 2 mM L-glutamine.

Quality Control:

• 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.
• These cells are guaranteed mycoplasma-free.

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 1 ml of blasticidin (10 mg/ml)
• 1 ml of puromycin (10 mg/ml)
• 2 x 1 ml of HEK-Blue™ CLR Selection (250X) 
• 1 ml of Normocin™ (50 mg/ml)
• 1 pouch of HEK-Blue™ Detection

Shipped on dry ice (Europe, USA, Canada and some areas in Asia)

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