CU-T12-9

Catalog No.S2979 Batch:S297901

Technical Data

Formula	C₁₇H₁₃F₃N₄O₂
Molecular Weight	362.31	CAS No.	1821387-73-8
Solubility (25°C)*	In vitro	DMSO	72 mg/mL (198.72 mM)
		Ethanol	5 mg/mL (13.8 mM)
		Water	Insoluble
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. * Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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Biological Activity

Description

CU-T12-9 is a specific agonist of Toll-like receptor (TLR) 1/2 with EC50 of 52.9 nM in HEK-Blue hTLR2 SEAP assay. CU-T12-9 activates both the innate and the adaptive immune systems. CU-T12-9 signals through NF-κB and invokes an elevation of the downstream effectors TNF-α, IL-10, and iNOS.

Targets

TLR 1/2 ^[1]
(HEK-Blue hTLR2 SEAP assay)

52.9 nM(EC50)

In vitro

CU-T12-9 specifically induces TLR1/2 activation, and facilitates the TLR1/2 heterodimeric complex formation, which in turn activates the downstream NF-κB signaling.^[1]

Protocol (from reference)

Cell Assay:^{^[1]}	Cell lines HEK-Blue hTLR2 cells Concentrations 60 nM Incubation Time 24 h Method Cells were cultured in 200 μl of DMEM supplemented with 10% FBS, 10× penicillin/streptomycin, and 10× l-glutamine. Cells were implanted in 96-well plates (4 × 10<sup>4</sup> cells per well) for 24 hours at 37°C before drug treatment. In the next 24 hours of treatment, medium was removed from the 96-well plate and replaced with 200 μl of supplemented Opti-MEM [0.5% FBS, penicillin (50 U/ml), streptomycin (50 μg/ml), 1× nonessential amino acids] containing 60 nM CU-T12-9, 0.66 nM (1 ng/ml) Pam3CSK4, or 0.77 nM (1 ng/ml) Pam2CSK4 according to the manufacturer’s protocol (InvivoGen), as well as different antibodies (0 to 10 μg/ml), including anti-hTLR1–IgG, anti-hTLR2–IgA, or anti-hTLR6–IgA. A sample buffer (20 μl) from each well of the cell culture supernatants was collected and transferred to a transparent 96-well plate. Each well was treated with 200 μl of QUANTI-Blue buffer and incubated at 37°C for 1 h.

Cell Assay:^{^[1]}

Cell lines
HEK-Blue hTLR2 cells
Concentrations
60 nM
Incubation Time
24 h
Method
Cells were cultured in 200 μl of DMEM supplemented with 10% FBS, 10× penicillin/streptomycin, and 10× l-glutamine. Cells were implanted in 96-well plates (4 × 10<sup>4</sup> cells per well) for 24 hours at 37°C before drug treatment. In the next 24 hours of treatment, medium was removed from the 96-well plate and replaced with 200 μl of supplemented Opti-MEM [0.5% FBS, penicillin (50 U/ml), streptomycin (50 μg/ml), 1× nonessential amino acids] containing 60 nM CU-T12-9, 0.66 nM (1 ng/ml) Pam3CSK4, or 0.77 nM (1 ng/ml) Pam2CSK4 according to the manufacturer’s protocol (InvivoGen), as well as different antibodies (0 to 10 μg/ml), including anti-hTLR1–IgG, anti-hTLR2–IgA, or anti-hTLR6–IgA. A sample buffer (20 μl) from each well of the cell culture supernatants was collected and transferred to a transparent 96-well plate. Each well was treated with 200 μl of QUANTI-Blue buffer and incubated at 37°C for 1 h.

References

[1] Kui Cheng, et al. Sci Adv. 2015;1(3):e1400139.

Selleck's CU-T12-9 has been cited by 2 publications

NHWD-1062 ameliorates inflammation and proliferation by the RIPK1/NF-κB/TLR1 axis in Psoriatic Keratinocytes [ Biomedicine & Pharmacotherapy, 2023, 114638]	PubMed: None
NHWD-1062 ameliorates inflammation and proliferation by the RIPK1/NF-κB/TLR1 axis in Psoriatic Keratinocytes [ Biomed Pharmacother, 2023, 162:114638]	PubMed: 37011486

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SHIPPING AND STORAGE
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