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HEK-Blue™ mMincle Cells

HEK-Blue™ mMincle Cells Unit size Cat. code Docs Qty Price
Murine Mincle / NF-κB / SEAP reporter HEK293 cells
3-7 x 10e6 cells
hkb-mmcl
+-
$1,182.00

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 enquiry.

 

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.

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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), 50 U/ml penicillin, 50 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.
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Contents

Shipped on dry ice (Europe, USA & Canada)

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Citations

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