HEK-Blue™ mDectin-2 Cells

SEAP Reporter 293 cells expressing the murine Dectin-2 gene

HEK-Blue™ mDectin-2 cells have been engineered to express a secreted alkaline phosphatase (SEAP) in response to stimulation by Dectin-2 ligands.

Dectin-2 is a C-type lectin receptor (CLR) that binds high mannose-type carbohydrates and has been demonstrated to be the functional receptor for α-mannans.  This receptor plays an important role in antifungal innate immunity [1, 2].

HEK-Blue™ mDectin-2 cells are derived from the human embryonic kidney HEK293 cells which do not naturally express Dectin-2. We have engineered HEK-Blue™ cells that stably express high levels of murine Dectin-2 as well as the genes of the Dectin-2 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™ mDectin-2 cells  reporter cell lines respond to Dectin-2 ligands, such as Furfurman. 

They do not respond to other CLR ligands, such as trehalose-6,6-dibehenate (TDB), a Mincle ligand.

To identify Dectin-2-specific responses, we recommend to use HEK-Blue™ Null1-v cells as a control cell line.

References:

1.  Hou H. et al, 2017. C-type Lectin Receptor: Old Friend and New Player. Med Chem. 2017;13(6):536-543. 
2. Wang HL. et al., 2017. Ligation of Dectin-2 with a novel microbial ligand promotes adjuvant activity for vaccination. PLoS Pathog. 213(8):e1006568.

Figures

HEK-Blue™ mDectin-2 and HEK-Blue™ Null1-v cells (parental cell line) were stimulated with various CLR agonists: zymosan depleted (100 µg/ml), zymosan (10 µg/ml), furfurman (10 µg/ml), HKCA (1x10⁸ cells), HKSC (1x10⁸ cells), TDB (10 µg/ml), and TNF-α (100 ng/ml). TNF-α, an NF-κB activator, was used as a positive control. NF-κB-induced SEAP activity was assessed using HEK-Blue™ Detection and by reading the optical density (OD) at 655 nm. Non-induced cells (NI) served as negative controls.

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™ mDectin-2 cells have been stimulated by various pathogen recognition receptor (PRR) agonists. As expected, Dectin-2 agonists induced the production of SEAP.
• The cell surface expression of murine Dectin-2 (mDectin-2) 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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