TLR3-IN-1 is a potent, highly selective TLR3 signaling inhibitor. TLR3-IN-1 represses the expression of downstream signaling pathways mediated by the TLR3/dsRNA complex, including TNF-α and IL-1β^[1].

TLR3-IN-1 shows dose-dependent inhibitory effects blocking Poly (I:C)-induced TLR3 activation with an IC₅₀ of 3.44 µM^[1].
TLR3-IN-1 competes with dsRNA for binding to TLR3 with a K_i of 2.96 µM^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

377.82

C₁₈H₁₃ClFNO₃S

1279713-77-7

Room temperature in continental US; may vary elsewhere.

Powder	-20°C	3 years
	4°C	2 years
In solvent	-80°C	6 months
	-20°C	1 month

In Vitro: 

DMSO : 100 mg/mL (264.68 mM; Need ultrasonic)

*

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

In Vivo:

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： 10% DMSO    40% PEG300    5% Tween-80    45% saline

  Solubility: ≥ 2.5 mg/mL (6.62 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (6.62 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: ≥ 2.5 mg/mL (6.62 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (6.62 mM，饱和度未知) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Yibo Wang, et al. Small-Molecule Modulators of Toll-like Receptors. Acc Chem Res. 2020 May 19;53(5):1046-1055.

RAW 264.7 cells are planted in 6-well plates in duplicate at 1,000,000 cells per well with 3 mL of medium (RPMI 1640 medium, supplemented with 10% FBS, Penicillin (100 U/mL) and Streptomycin (100 mg/mL)) and grown for 24 h at 37°C in a 5% CO₂ humidified incubator. After 24 h, non-adherent cells and media are removed and replaced with fresh RPMI 1640 medium (3 mL/well). Two wells of adherent macrophages are treated with Poly (I:C) (15 μg/mL). One of the two wells is treated with 27 μM CU CPT 4a. One additional well is treated with only CU CPT 4a (27 μM) only. Plates are then incubated for an additional 24 h. After removal of the medium, cells are washed with PBS (3×1 mL), the 6 well plate is put on ice, then 500 μL of lysis buffer is added in each well (Lysis Buffer: 120 μL 0.5M EDTA; 12 mL Mammalian Protein Extraction Reagent, 100 μL cocktail, 0.36 mL NaCl (5 M, aqueous) ). After 5 min, the mixture is transferred into corresponding 1.5 mL tube, spun for 20 min at 13.2 K rpm in a cold room. Approximately 400 μL of supernatant are collected into new tubes, frozen at -80°C until ready for cytokine measurement^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Yibo Wang, et al. Small-Molecule Modulators of Toll-like Receptors. Acc Chem Res. 2020 May 19;53(5):1046-1055.