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HEK-Blue™ hDectin-1b Cells

HEK-Blue™ hDectin-1b cells Unit size Cat. code Docs Qty Price
human Dectin-1b-expressing HEK 293 cells
3-7 x 10e6 cells
hkb-hdect1b
+-
$1,142.00

SEAP Reporter 293 cells expressing the human Dectin-1b gene

HEK-Blue™ hDectin-1b cells have been engineered to express a secreted alkaline phosphatase (SEAP) in response to stimulation by Dectin-1 ligands

Dectin-1 is a C-type lectin receptor (CLR) that recognizes β-glucans, glucose polymers found in the cell walls of fungi, and thus plays an important role in antifungal innate immunity. Dectin-1 is alternatively spliced into 2 major isoforms: a full-length A isoform and a ‘stalkless’ B isoform, which do not induce the same response to soluble and particulate β-glucans [1].

HEK-Blue™ hDectin-1b cells are derived from the human embryonic kidney HEK293 cells which do not naturally express Dectin-1. We have engineered HEK-Blue™ cells that stably express high levels of human Dectin-1b isoform and genes involved in the Dectin-1/NF-κB/SEAP signaling pathway. Levels of SEAP are readily measurable in real-time by a simple colorimetric assay with HEK-Blue™ Detection or QUANTI-Blue™ . In contrast to cells expressing the human Dectin‑1a gene, HEK‑Blue™ hDectin‑1b cells do not respond to soluble β-glucans, such as laminarin and WGP soluble.

Read our review on Dectin-1 & β-glucans.

Read our review on Dectin-1 & inflammasomes.

 

1. Heinsbroek, S.E, et al, 2006. Expression of functionally different Dectin-1 isoforms by murine macrophages. J Immunol. 176: 5513.

Evaluation of NF-κB responses to Dectin-1 ligands
Evaluation of NF-κB responses to Dectin-1 ligands

NF-κB responses of HEK-Blue™ hDectin-1a and -1b and HEK-Blue™ Null I-v cells (control cell line) to Dectin-1 ligands. Cells were incubated with particulate ligands such as Zymozan (10 μg/ml), WGP dispersible (100 μg/ml) and HKCA (3 x106 cells/ml), or soluble ligands such as Laminarin (100 μg/ml), WGP soluble (10 μg/ml) or TDB (10 μg/ml). TNF-α (10 ng/ml) was used as a positive control. After 24h, SEAP activity was assessed in the supernatant using QUANTI-Blue™, by reading the optical density (OD) at 630 nm.

Inhibition of HEK-Blue™ hDectin-1b responses by Laminarin and WGP soluble
Inhibition of HEK-Blue™ hDectin-1b responses by Laminarin and WGP soluble

Dose-dependent inhibition HEK-Blue™ hDectin-1b responses by Laminarin and WGP soluble. Cells were incubated with particulate ligands such as Zymozan (30 μg/ml) or HKCA (3 x106 cells/ml) and soluble ligands such as (a) Laminarin (starting concentration 100 μg/ml) or (b) WGP soluble (starting concentration 1 mg/ml). After 24h, SEAP activity was assessed in the supernatant using QUANTI-Blue™. Data are presented as the percentage of SEAP activity measured in presence of Zymosan or HKCA without Laminarin or WGP soluble.

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Specifications

Required Selective Antibiotics: 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.

Guaranteed mycoplasma-free.

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Contents

Shipped on dry ice

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Details

Quality Control
HEK-Blue™ hDectin-1b cells have been stimulated by various pathogen recognition receptor (PRR) agonists. As expected, Dectin-1 agonists induced the production of SEAP. The cell surface expression of human Dectin-1b (hDectin-1b) 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™ hDectin-1b cells:
As their parental cell line, HEK293 cells, express endogenous levels of TLR3, TLR5 and NOD1, HEK-Blue™ hDectin-1b cells will respond to TLR3, TLR5 and NOD1 agonists, such as poly(I:C), flagellin and C12‑iE‑DAP, respectively.
In order to identify Dectin-1-specific responses, we recommend to use HEK-Blue™ Null1-v cells as a control cell line.

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